Triazole derivatives as Wnt signaling pathway inhibitors

ABSTRACT

The present invention relates to compounds of formula I, to processes for their preparation, to pharmaceutical formulations containing such compounds and to their use in therapy: 
     
       
         
         
             
             
         
       
     
     Such compounds find particular use in the treatment and/or prevention of conditions or diseases which are affected by over-activation of signaling in the Wnt pathway and increased presence of nuclear β-catenin. For example, these may be used in preventing and/or retarding proliferation of tumor cells and metastasis, for example carcinomas such as colon carcinomas.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/992,879 filed on Oct. 21, 2013, which is a §371 of PCT/GB2011/052441filed on Dec. 8, 2011, which claims the benefit of priority to U.S.Provisional Application Ser. No. 61/420,942 filed on Dec. 8, 2010, whichare incorporated herein by reference.

The present invention relates to compounds, to pharmaceuticalformulations containing such compounds and to their use in therapy, inparticular as Wnt signaling pathway inhibitors for reducing theproliferation of tumor cells and metastasis. The invention furtherrelates to processes for the preparation of such compounds and tointermediates formed during these processes.

The Wnt family of glycoproteins control a variety of developmentalprocesses including cell fate specification, proliferation, polarity andmigration. Consequently, the Wnt pathway is instrumental in ensuringproper tissue development in embryos and tissue maintenance in adults.There are at least three signaling pathways involved in the Wnt signaltransduction process. The canonical (or β-catenin dependent) Wnt pathwaywas discovered first and has been studied most. In the absence of a Wntsignal, the transcriptional activator β-catenin is a phosphorylatedintracellular multi-protein complex which is subsequently degraded.Within this complex the AXIN and adenomatous polyposis coli (APC)proteins form a scaffold that facilitates β-catenin phosphorylation bycasein-kinase1α (CK1α) and glycogen synthase kinase 3β (GSK-3β).Phosphorylated β-catenin is subsequently ubiquitinylated, resulting inits degradation in the proteasome. When Wnt signaling is inactive andtherefore levels of free β-catenin are low, DNA-binding T-cellfactor/lymphoid enhancer factor (TCF/LEF) proteins interact withtranscriptional repressors to block Wnt target gene expression in thenucleus. Binding of Wnt molecules to FZD-LRP receptor complexes at themembrane leads to a cascade of events that lead to the inactivation ofthe β-catenin destruction complex. This allows β-catenin to accumulateand enter the nucleus where it interacts with members of the Tcf/Leffamily and converts the Tcf proteins into potent transcriptionalactivators by recruiting co-activator proteins ensuring efficientactivation of Wnt target genes.

Canonical Wnt signaling is over-activated in a variety of tumors whereit plays a central role in cell growth and tumor progression (Barker etal., Nat. Rev. Drug. Discov. 5: 997-1014, 2006; Grigoryan et al., GenesDev. 22: 2308-2341, 2008; and Shitashige et al., Cancer Sci. 99:631-637, 2008). About 90% of sporadic colon cancers show aberrant Wntsignaling (Liu et al., Nat. Genet. 26: 146-147, 2000; and Morin et al.,Science 275: 1787-1790, 1997), while all pancreatic adenocarcinomasexhibit alterations in Wnt/Notch signaling (Jones et al., Science 321:1801-1806, 2008).

Wnt activating mutations are present in a variety of cancers includinggastric cancer, hepatocellular carcinoma, Wilms tumor of the kidney,medulloblastoma, melanoma, non-small cell lung cancer, ovarianendometriod cancer, anaplastic thyroid cancer, pancreas adenocarcinoma,and prostate cancer (Barker et al. supra). Mutations in the adenomatouspolyposis coli gene (APC), β-catenin, or Axin genes lead to accumulationof nuclear β-catenin and such mutations are frequently associated withcolon cancer (Morin et al. supra). Furthermore, alterations inextracellular proteins which silence Wnt signaling including secretedfrizzled related proteins (SFRPs) (Suzuki et al., Nat. Genet 36:417-422, 2004), Dickkopf (Dkk) (Aguilera et al., Oncogene 25: 4116-4121,2006) and members of the Wnt inhibitor factor (WIF) family (Mazieres etal., Cancer Res. 64: 4717-4720, 2004) can also lead to abnormal pathwayactivity (Polakis, Curr. Opin. Genet. Dev. 17: 45-51, 2007).

Blocking canonical Wnt activity in colorectal and other Wnt deregulatedcancers has been shown to cause cell cycle arrest in G1 and this is acrucial step in inhibiting tumor cell growth (van de Wetering et al.,Cell 111: 241-250, 2002; and Sukhdeo et al., Proc. Natl. Acad. Sci. USA104: 7516-7521, 2007). In recent years, several classes ofsmall-molecules have been shown to act as Wnt inhibitors. These drugsexert their inhibitory effects at various levels of the Wnt signalingpathway. Small molecules, interfering with nuclear TCF/β-catenin bindingand with the cyclic AMP response element-binding protein (CBP), havebeen identified and described (Emami et al., Proc. Natl. Acad. Sci. USA101: 12682-12687, 2004; and Lepourcelet M et al., Cancer Cell 5: 91-102,2004). Topo IIα and PARP-1 (Shitashige et al., Cancer Sci. 99: 631-637,2008) or TBP, BRG1, BCL9, pygopus and Hyrax (Barker et al. supra) havebeen proposed to be potential targets for inhibiting canonical Wntsignaling. Recently, two groups of chemical substances (IWR-1 andXAV939) have been identified which stabilize the destruction complex(Chen et al., Nat. Chem. Biol. 5: 100-107, 2009; and Huang et al.,Nature: 461: 614-620, 2009). By blocking the PARP domain of Tankyrase,XAV939 and IWR-1 are thought to alter the PARsylation and ubiquitinationof AXIN2 that results in its increased stability and in inhibition ofcanonical Wnt signaling. Since elevated levels of β-catenin in thenucleus are a common feature of abnormal canonical Wnt signaling,down-regulation of canonical Wnt activity by reducing the presence ofβ-catenin represents a potential therapeutic strategy.

We have now found a selected class of compounds which exhibit anactivity in blocking canonical Wnt signaling, and in particular whichare capable of reducing levels of activated nuclear β-catenin. To theextent that these are able to affect the stability of activatedβ-catenin downstream of APC and GSK-3β, these are considered to offerbroader potential than other compounds known to act further upstream inthe canonical Wnt signaling pathway. Such compounds are suitable forinhibiting the proliferation of tumor cells in general and, inparticular, those associated with colorectal cancers, breast cancer,non-small cell lung cancer, prostate cancer and pancreaticadenocarcinoma.

The invention provides compounds of general formula I:

(wherein

-   Z¹ represents an unsaturated, 5- to 7-membered heterocyclic ring, or    -   a group of the formula —(CH₂)_(x)—CO—NH—NH—CO— (CH₂)_(y)—        wherein x and y independently denote an integer from 0 to 2;-   Z² represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(a);    -   where each R_(a) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl,        C₁₋₄haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —C(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR,        —S(O)R, —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkenyl or C₂₋₆        alkynyl);-   R¹ represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(b);    -   where each R_(b) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl) optionally interrupted by one or more        —O—, —S— or —NR— groups (preferably by one or two —O— atoms),        C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl (e.g. CF₃), —CN,        —NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂, —C(O)NR₂,        —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R, —S(O)₂R, —S(O)OR or        —S(O)₂NR₂ group (where each R is independently H, C₁₋₆ alkyl,        C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl);-   R² represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(c);    -   where each R_(c) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R,        —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or        C₂₋₆ alkynyl);-   L¹ represents a C₁₋₄ alkylene group optionally substituted by one or    more (e.g. 1 or 2) groups R_(d), wherein one or more (preferably one    or two) methylene groups are each replaced by a group selected from    —CR_(e)═CR_(f)—, and —C═C═C—; and wherein one or more (preferably    one to three) methylene groups may each additionally be replaced by    a group Y¹;    -   where each Y¹ is independently selected from —O—, —S—, —NH—,        —NR′″—, —NR′″—C(O)—, —C(O)—NR′″—, —C(O)—, —S(O₂)—, —S(O)— and        —CR′″═N— (where each R′″ is independently hydrogen or C₁₋₆        alkyl);    -   where each R_(d) may be identical or different and may be        selected from C₁₋₆ alkyl (preferably C₁₋₃ alkyl), hydroxy, C₁₋₆        alkoxy (e.g. C₁₋₃ alkoxy) and halogen (i.e. F, Cl, Br and I,        preferably F); and    -   where R_(e) and R_(f) are independently selected from H, C₁₋₃        alkyl, halogen (e.g. F or Cl), C₁₋₃ haloalkyl (e.g. —CF₃), —CN,        —NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OPO₃R, —OSO₂R and        —OSiR₄ (where each R is independently H, C₁₋₆ alkyl or C₁₋₆        haloalkyl);-   L² represents a bond or an optionally substituted C₁₋₆ alkylene    group; and-   L³ represents a bond or an optionally substituted C₁₋₆ alkylene    group)-   the isomers (e.g. stereoisomers), pharmaceutically acceptable salts,    and prodrugs thereof.

In any of the embodiments of the invention herein described, it isenvisaged that in the definition of the linker L¹ all methylene groupsmay be replaced by a group —CR_(e)═CR_(f)—, —C≡C— or —C═C═C—). Suitablelinkers, L¹, therefore include groups which contain no methylene moiety(i.e. where all such groups have been replaced by a group—CR_(e)═CR_(f)—, —C≡C— or —C═C═C—).

In formula I, L³ is preferably a direct bond or a —CH₂— group.

Preferred compounds in accordance with the invention are those ofgeneral formula II:

(wherein

-   Z¹ represents an unsaturated, 5- to 7-membered heterocyclic ring, or    -   a group of the formula — (CH₂)_(x)—CO—NH—NH—CO— (CH₂)_(y)—        wherein x and y independently denote an integer from 0 to 2;-   Z² represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(a);    -   where each R_(a) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R,        —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or        C₂₋₆ alkynyl);-   R¹ represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(b);    -   where each R_(b) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R, —S(O)₂R        or —S(O)OR group (where each R is independently H, C₁₋₆ alkyl,        C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl);-   R² represents an aryl or heteroaryl group (preferably an aryl group)    optionally substituted by one or more (e.g. 1, 2, 3 or 4) groups    R_(c);    -   where each R_(c) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R,        —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or        C₂₋₆ alkynyl);-   L¹ represents a C₁₋₄ alkylene group optionally substituted by one or    more (e.g. 1 or 2) groups R_(d), wherein one or more (preferably one    or two) methylene groups are each replaced by a group selected from    —CR_(e)═CR_(f)—, —C≡C— and —C═C═C—; and    -   wherein one or more (preferably one to three) methylene groups        may each additionally be replaced by a group Y¹;        -   where each Y¹ is independently selected from —O—, —S—, —NH—,            —NR′″—, —NR′″—C(O)—, —C(O)—NR′″—, —C(O)—, —S(O₂)—, —S(O)—            and —CR′″═N— (where each R′″ is independently hydrogen or            C₁₋₆ alkyl);        -   where each R_(d) may be identical or different and may be            selected from C₁₋₆ alkyl (preferably C₁₋₃ alkyl), hydroxy,            C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy) and halogen (i.e. F, Cl, Br            and 1, preferably F); and        -   where R_(e) and R_(f) are independently selected from H,            C₁₋₃ alkyl, halogen (e.g. F or Cl), C₁₋₃ haloalkyl (e.g.            —CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R,            —OPO₃R, —OSO₂R and —OSiR₄ (where each R is independently H,            C₁₋₆ alkyl or C₁₋₆ haloalkyl);-   L² represents a bond or an optionally substituted C₁₋₆ alkylene    group, preferably a bond)-   the isomers (e.g. stereoisomers), pharmaceutically acceptable salts,    and prodrugs thereof.

In the compounds herein described, preferred linkers L¹ comprise a C₁₋₄alkylene group (preferably a C₁₋₂ alkylene) optionally substituted byone or more (e.g. 1 or 2) groups R_(d), wherein one or more (preferablyone or two) methylene groups are each replaced by a group—CR_(e)═CR_(f)— or by a group —C≡C—;

-   -   where each R_(d) may be identical or different and may be        selected from C₁₋₆ alkyl (preferably C₁₋₃ alkyl), hydroxy, C₁₋₆        alkoxy (e.g. C₁₋₃ alkoxy) and halogen (i.e. F, Cl, Br and I,        preferably F); and    -   where R_(e) and R_(f) are independently selected from H, C₁₋₃        alkyl, halogen (e.g. F or Cl) and —CN.

In the compounds of formula I and II, where Z¹ is a group of the formula— (CH₂)_(x)—CO—NH—NH—CO— (CH₂)_(y)—, it is preferred that x and y areboth zero.

Particularly preferred compounds in accordance with the invention arethose of formula I or II wherein:

-   Z¹ represents a 5-membered heterocyclic ring containing two or three    heteroatoms selected from N, O and S;-   Z² represents phenyl, pyridyl, pyrimidinyl or oxadiazolyl optionally    substituted by one or more (e.g. 1, 2, 3 or 4) groups R_(a);    -   where each R_(a) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R,        —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or        C₂₋₆ alkynyl);-   R¹ represents an aryl or heteroaryl group optionally substituted by    one or more (e.g. 1, 2, 3 or 4) groups R_(b);    -   where each R_(b) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R, —S(O)₂R        or —S(O)OR group (where each R is independently H, C₁₋₆ alkyl,        C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl);-   R² represents an aryl group optionally substituted by one or more    (e.g. 1, 2, 3 or 4) groups R_(c);    -   where each R_(c) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl), C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄        haloalkyl (e.g. CF₃), —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR,        —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R,        —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R is        independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or        C₂₋₆ alkynyl);-   L¹ represents a C₁₋₄ alkylene group (preferably a C₁₋₂ alkylene)    optionally substituted by one or more (e.g. 1 or 2) groups R_(d),    wherein one or two methylene groups (e.g. one methylene group) are    each replaced by a group —CR_(e)═CR_(f)— or by a group —C≡C—;    -   where each R_(d) may be identical or different and may be        selected from C₁₋₆ alkyl (preferably C₁₋₃ alkyl), hydroxy and        C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy);    -   where R_(e) and R_(f) are independently selected from H, C₁₋₃        alkyl (e.g. methyl), halogen (e.g. F or Cl) and —CN; and-   L² represents a bond or an optionally substituted C₁₋₄ alkylene    group, preferably a bond)-   the isomers (e.g. stereoisomers), pharmaceutically acceptable salts    and prodrugs thereof.

More particularly preferred compounds according to the invention arethose of formula I or II wherein:

-   Z¹ represents a 5-membered heterocyclic ring containing two nitrogen    atoms and one oxygen atom;-   Z² represents phenyl, pyridyl or pyrimidinyl optionally mono- or    di-substituted by a group R_(a);    -   where R_(a) may be selected from halogen (i.e. F, Cl, Br, I),        hydroxy, C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy) and —S(O)₂R (where R is        H or C₁₋₃ alkyl);-   R¹ represents an aryl or heteroaryl group optionally    mono-substituted by group R_(b);    -   where R_(b) is selected from halogen (i.e. F, Cl, Br, I), C₁₋₆        alkyl (preferably C₁₋₃ alkyl), C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy),        hydroxy and —CN;-   R² represents an aryl group optionally mono- or di-substituted by a    group R_(c);    -   where each R_(c) may be identical or different and may be        selected from halogen (i.e. F, Cl, Br, I), C₁₋₆ alkyl        (preferably C₁₋₃ alkyl) and C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy);-   L¹ represents a C₁₋₄ alkylene group (preferably a C₁₋₂ alkylene)    optionally substituted by one or more (e.g. 1 or 2) groups R_(d),    wherein one or two methylene groups (e.g. one methylene group) are    each replaced by a group —CR_(e)═CR_(f)— or by a group —C≡C—;    -   where each R_(d) may be identical or different and may be        selected from C₁₋₆ alkyl (preferably C₁₋₃ alkyl), hydroxy and        C₁₋₆ alkoxy (e.g. C₁₋₃ alkoxy);    -   where R_(e) and R_(f) are independently selected from H, C₁₋₃        alkyl and halogen (e.g. F or Cl); and-   L² represents a bond, or a C₁₋₂ alkylene group)-   the isomers (e.g. stereoisomers), pharmaceutically acceptable salts    and prodrugs thereof.

Examples of group Z¹ in formula I and II include the following:

Of these structures, the following are particularly preferred for Z¹:

In a yet further preferred aspect the invention thus provides thefollowing compounds of formulae Ia, IIa, Ib and IIb:

(wherein Z², R¹, R², L¹, L² and L³ are as hereinbefore defined); theisomers (e.g. stereoisomers), pharmaceutically acceptable salts andprodrugs thereof.

In preferred embodiments, Z² represents an optionally substituted arylor heteroaryl group, preferably a phenyl, pyridyl or pyrimidinyl groupoptionally substituted by one or two (preferably one) groups R_(a) inwhich each R_(a) is independently halogen (preferably Cl or F), hydroxy,C₁₋₆ alkoxy (preferably C₁₋₃ alkoxy, e.g. methoxy) or —S(O)₂R (where Ris H or C₁₋₃ alkyl). Particularly preferably, Z² represents anoptionally substituted phenyl or pyridyl group, e.g. optionallysubstituted pyridyl. When substituted, the substituents on the ring mayindependently be selected from the group consisting of hydroxy, methoxy,ethoxy, chloro, fluoro and methylsulphonyl. Preferred substituents areethoxy, chloro, fluoro and methylsulphonyl. One or more of such groupsmay be present on the ring and in any ring position. However, it ispreferred that one or two such groups will be present. Where twosubstituents are present these will generally be identical.

Examples of group Z² include the following:

Particularly preferred groups Z² include the following:

In preferred embodiments, R¹ represents phenyl or pyridyl optionallysubstituted by one or two (preferably one) groups R_(b) in which eachR_(b) is independently halogen (e.g. F, Cl or Br), hydroxy, C₁₋₆ alkyl(preferably C₁₋₃ alkyl, e.g. methyl), C₁₋₆ alkoxy (preferably C₁₋₃alkoxy, e.g. methoxy) or cyano. Particularly preferably, R¹ represents asubstituted (e.g. mono-substituted) pyridyl ring. Preferred substituentsinclude hydroxy, methoxy and cyano groups, especially methoxy.

Examples of group R¹ include the following:

Particularly preferred groups R¹ include the following:

In preferred embodiments, R² represents phenyl or pyridyl optionallysubstituted by one or two groups R_(c) in which each R_(c) isindependently halogen (e.g. F or Cl) or C₁₋₆ alkoxy (preferably C₁₋₃alkoxy, e.g. methoxy or ethoxy).

Particularly preferably, R² is optionally substituted phenyl. Whensubstituted, the ring substituents on the phenyl group may independentlybe selected from the group consisting of C₁₋₃ alkyl (e.g. methyl orethyl), methoxy, ethoxy, chloro and fluoro. One or more of such groupsmay be present on the ring and in any ring position. However, it ispreferred that one or two such groups will be present. Particularlypreferably, the phenyl ring will be substituted by a single chloro groupeither in the ortho or para-position, e.g. in the ortho-position.

Examples of group R² include the following:

Particularly preferably, R² is a group:

In preferred embodiments, L¹ is a C₁ alkylene linker in which the singlemethylene group is either replaced by a group —CR_(e)═CR_(f)— (whereR_(e) and R_(f) are as hereinbefore defined), by a group —C≡C— or by agroup —C═C═C—.

Particularly preferred as linker groups L¹ are groups of the formula—CR_(e)═CR_(f)— in which R_(e) and R_(f) are independently selected fromH and C₁₋₃ alkyl. Such groups may be either cis or trans, althoughpreferably they will be in the trans configuration.

Particularly preferably, R_(e) and R_(f) in the linker group L¹ areidentical and are either both H or both methyl. A preferred linker groupL¹ is —CH═CH—.

Examples of L¹ include the following:

In preferred embodiments L² is a bond or a C₁₋₂ alkylene group (e.g.methylene). Preferably L² is a bond.

In preferred embodiments, L³ is a bond.

Particularly preferred compounds according to the invention are thefollowing compounds of formulae IIc and IId:

(wherein

-   Z² is an optionally substituted pyridyl, phenyl or pyrimidinyl ring,    preferably a phenyl ring substituted by one or two (preferably by    one) halo atoms (e.g. Cl or F) or by one methylsulphonyl group, or a    pyridyl ring optionally substituted by a halo atom (e.g. Cl), by an    alkoxy group (e.g. ethoxy) or by a methylsulphonyl group;-   R¹ is a substituted phenyl or pyridyl ring, preferably a phenyl or    pyridyl ring substituted by a halo atom (e.g. Cl), a C₁₋₆ alkoxy    (e.g. methoxy) group, a hydroxy or cyano group;-   R² is an optionally substituted phenyl ring, preferably a phenyl    ring substituted by one halo atom (e.g. CI);-   L¹ is cis or trans —CH═CH—, preferably trans —CH═CH—)-   the isomers (e.g. stereoisomers), pharmaceutically acceptable salts    and prodrugs thereof.

The following are examples of particularly preferred compounds inaccordance with the invention:

Compound No. Structure  (1)

 (2)

 (3)

 (4)

 (5)

 (6)

 (7)

 (8)

 (9)

(10)

(11)

(12)

(13)

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(58)

Particularly preferred compounds in accordance with the invention areCompound Nos. (1), (2), (3), (4), (5), (6), (7), (8), (10), (11), (12),(14), (33), (56), (57) and (58), their isomers, pharmaceuticallyacceptable salts thereof and prodrugs. More particularly preferredcompounds in accordance with the invention are Compound Nos. (2), (3),(4), (8), (10), (11), (12), (14) and (57), their isomers,pharmaceutically acceptable salts and prodrugs.

In one embodiment the following compounds per se are excluded from thescope of the invention:

Unless otherwise stated, all substituents are independent of oneanother.

In the case where a subscript is the integer 0 (i.e. zero), it isintended that the group to which the subscript refers is absent, i.e.there is a direct bond between the groups either side of that particulargroup.

Unless otherwise stated, any reference herein to a “bond” is intended torefer to a saturated bond.

In the case where an asterisk (*) is present in any of the structuralformulae of any of the substituents provided herein, this is to beunderstood as indicating the point of attachment of that substituent tothe remainder of the molecule. Where any of these formulae include twoasterisks (denoting two points of attachment), either one of these maybe linked to a desired point of attachment on the remainder of themolecule. The orientation of such structures specifically presentedherein is not intended to imply that these must be linked in theorientation which is given.

Unless otherwise stated, the term “halo” or “halogen atom” may befluoro, chloro, bromo, or iodo. Preferably, this is fluoro or chloro.

As used herein, the term “alkyl” refers to a saturated hydrocarbon groupand is intended to cover both straight-chained and branched alkylgroups. Examples of such groups include methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl,iso-pentyl, neo-pentyl, n-hexyl, etc. An alkyl group preferably containsfrom 1-6 carbon atoms, e.g. 1-4 carbon atoms. Unless otherwise stated,any alkyl group mentioned herein may optionally be substituted by one ormore groups, which may be identical or different, for example hydroxy,alkoxy, acyloxy, amino or halogen atoms (e.g. F, Cl or Br).

As used herein, the term “alkenyl” refers to an alkyl group having oneor more carbon-carbon double bonds and includes both straight-chainedand branched alkenyl groups. The term “C₂₋₆ alkenyl” refers to analkenyl group having from 2 to 6 carbon atoms and one or more (e.g. oneor two) double bonds. Examples of such groups include vinyl, allyl,propenyl, iso-propenyl, butenyl, iso-butenyl, crotyl, pentenyl andhexenyl. Unless otherwise stated, any alkenyl group mentioned herein mayoptionally be substituted by one or more groups, which may be identicalor different, for example hydroxy, alkoxy, acyloxy, amino or halogenatoms (e.g. F, Cl or Br).

As used herein, the term “alkynyl” refers to an alkyl group having oneor more carbon-carbon triple bonds and includes both straight-chainedand branched alkynyl groups. Unless otherwise stated, any alkynyl groupmentioned herein may optionally be substituted by one or more groups,which may be identical or different, for example hydroxy, alkoxy,acyloxy, amino or halogen atoms (e.g. F, Cl or Br).

As used herein, the term “haloalkyl” refers to an alkyl group having oneor more halo substituents. Examples of such groups include —CH₂F, —CHF₂,—CF₃, —CCl₃, —CHCl₂, —CH₂CF₃, etc.

As used herein, the term “alkylene” refers to a linking alkyl group andis intended to cover any straight-chained or branched alkylene group.Examples of such groups include methylene, ethylene, ethane-1,1-diyl,propylene, propane-2,2-diyl, 1-methylethylene, butylene,1-methylpropylene, 1,1-dimethyl ethylene, 1,2-dimethyl ethylene, etc.

As used herein, the term “unsaturated heterocyclic ring” is intended tocover any 5-, 6- or 7-membered, mono-, di or tri-unsaturatedheterocyclic ring which contains at least one heteroatom selected fromnitrogen, oxygen and sulphur. The heterocyclic ring structure may belinked to the remainder of the molecule through a carbon atom or, ifpresent, through a nitrogen atom. For example, it may be linked throughtwo carbon atoms, through two nitrogen atoms, or through one carbon andone nitrogen atom. Preferably it will be linked to the remainder of themolecule through two carbon atoms. Unless otherwise stated, anyheterocyclic ring mentioned herein may optionally be substituted by oneor more groups, which may be identical or different, for examplehydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino, cyano, nitro or halogen atoms(e.g. F, Cl or Br). A heterocyclic ring may further contain one or morecarbonyl or thiocarbonyl functionalities such that this includes oxo andthio-systems.

Illustrative examples of “unsaturated heterocyclic rings” are theheterocycles pyrrole, 2H-pyrrole, furan, pyrroline, thiophene, pyrazole,imidazole, oxazole, isoxazole, pyrazoline, imidazoline, thiazole,isothiazole, thiadiazole, pyridine, 2H-pyran, 4H-pyran, pyridazine,pyrimidine, pyrazine, 1,3-dioxine, 1,4-dioxine and triazole. Of these,thiazole, thiadiazole, pyrimidine, pyridazine, pyrazole, thiophene andtriazole are particularly preferred.

As used herein, the term “aryl” is intended to cover aromatic ringsystems. Such ring systems may be monocyclic or polycyclic (e.g.bicyclic) and contain at least one unsaturated aromatic ring. Wherethese contain polycyclic rings, these may be fused. Preferably suchsystems contain from 6-20 carbon atoms, e.g. either 6 or 10 carbonatoms. Examples of such groups include phenyl, 1-napthyl and 2-napthyl.A preferred aryl group is phenyl. Unless stated otherwise, any “aryl”group may be substituted by one or more substituents, which may beidentical or different, for example C₁ alkyl groups, hydroxy, methoxy,trifluoromethoxy and halo groups.

As used herein, the term “heteroaryl” is intended to cover heterocyclicaromatic groups. Such groups may be monocyclic or bicyclic and containat least one unsaturated hetero aromatic ring system. Where these aremonocyclic, these comprise 5- or 6-membered rings which contain at leastone heteroatom selected from nitrogen, oxygen and sulphur and containsufficient conjugated bonds to form an aromatic system. Where these arebicyclic, these may contain from 9-11 ring atoms. Examples of heteroarylgroups include thiophene, thienyl, pyridyl, thiazolyl, furyl, pyrrolyl,triazolyl, imidazolyl, oxadiazolyl, oxazolyl, pyrazolyl, imidazolonyl,oxazolonyl, thiazolonyl, tetrazolyl, thiadiazolyl, benzimidazolyl,benzooxazolyl, benzofuryl, indolyl, isoindolyl, pyridonyl, pyridazinyl,pyrimidinyl, imidazopyridyl, oxazopyridyl, thiazolopyridyl,imidazopyridazinyl, oxazolopyridazinyl, thiazolopyridazinyl and purinyl.Unless stated otherwise, any “heteroaryl” may be substituted by one ormore substituents, which may be identical or different, for example C₁₋₄alkyl groups, hydroxy, methoxy, trifluoromethoxy and halo groups.

The term “prodrug” is intended to encompass any compound which underphysiological conditions is converted into any of the compounds hereindescribed, i.e. a compound of formula I, II, Ia, IIa, Ib, IIb, IIc orIId. Suitable prodrugs include compounds which are hydrolysed underphysiological conditions to the desired molecule.

The compounds according to the invention may be prepared from readilyavailable starting materials using synthetic methods known in the art.Preferably, the compounds are obtained in accordance with the followingmethods which form part of the invention:

(a) reacting a compound of general formula III:

(wherein Z², R² and L³ are as hereinbefore defined)with a compound of general formula IV:

(wherein R¹, Z¹ and L¹ are as hereinbefore defined and L denotes aleaving group such as a halogen atom, e.g. Cl).

The reaction is conveniently carried out in a solvent or mixture ofsolvents, such as for example a polar solvent such as acetonitirile,acetone, DMF, DMSO or dioxane, in the presence of a dehydrating agentsuch as phosphoryl chloride, expediently at temperatures up to 150° C.,preferably at temperatures between −20 and 80° C.

(b) reacting a compound of general formula III:

(wherein Z², R² and L³ are as hereinbefore defined)with a compound of general formula V:

(wherein R¹ and L′ are as hereinbefore defined).

The reaction is conveniently carried out in a solvent or mixture ofsolvents, such as for example a polar solvent such as acetonitirile,acetone, DMF, DMSO or dioxane, in the presence of a dehydrating agentsuch as phosphoryl chloride, expediently at temperatures up to 150° C.,preferably at temperatures between −20 and 80° C.

(c) if desired, resolving a compound thus obtained into thestereoisomers thereof; and/or

(d) converting a compound thus obtained into a salt thereof,particularly a pharmaceutically acceptable salt thereof.

The compounds used as starting materials are either known from theliterature or may be commercially available. Alternatively, these may beobtained by methods known from the literature.

The invention includes all optical isomers and stereoisomers of thecompounds herein disclosed. In particular, the invention extends to theenantiomers of any of the compounds having a chiral centre in the groupL¹.

The compounds of general formulae I, II, Ia, IIa, Ib, IIb, IIc or IIdmay be resolved into their enantiomers and/or diastereomers. Forexample, where these contain only one chiral centre, these may beprovided in the form of a racemate or may be provided as pureenantiomers, i.e. in the R- or S-form. Any of the compounds which occuras racemates may be separated into their enantiomers by methods known inthe art, such as column separation on chiral phases or byrecrystallisation from an optically active solvent. Those compounds withat least two asymmetric carbon atoms may be resolved into theirdiastereomers on the basis of their physical-chemical differences usingmethods known per se, e.g. by chromatography and/or fractionalcrystallisation, and where these compounds are obtained in racemic form,they may subsequently be resolved into the enantiomers.

The invention further includes certain structural isomers of thecompounds herein disclosed. In particular, the invention extends totautomers of any of the compounds. As will be appreciated, certaincompounds according to the invention may exist in tautomeric forms, i.e.in forms which readily interconvert by way of a chemical reaction whichmay involve the migration of a proton accompanied by a switch of asingle bond and adjacent double bond. In cases where one of the groupsR_(e) and R_(f) is hydroxyl the compounds of the invention may, inparticular, undergo keto-enol tautomerism. Dependent on the conditions,the compounds may predominantly exist either in the keto or enol formand the invention is not intended to be limited to the particular formshown in any of the structural formulae given herein.

The compounds according to the invention may be converted into a saltthereof, particularly into a pharmaceutically acceptable salt thereofwith an inorganic or organic acid or base. Acids which may be used forthis purpose include hydrochloric acid, hydrobromic acid, sulphuricacid, sulphonic acid, methanesulphonic acid, phosphoric acid, fumaricacid, succinic acid, lactic acid, citric acid, tartaric acid, maleicacid, acetic acid, trifluoroacetic acid and ascorbic acid. Bases whichmay be suitable for this purpose include alkali and alkaline earth metalhydroxides, e.g. sodium hydroxide, potassium hydroxide or cesiumhydroxide, ammonia and organic amines such as diethylamine,triethylamine, ethanolamine, diethanolamine, cyclohexylamine anddicyclohexylamine. Procedures for salt formation are conventional in theart.

In a further aspect there is provided pharmaceutical formulationscomprising a compound of formula I, II, Ia, IIa, Ib, IIb, IIc or IId asherein defined, or a pharmaceutically acceptable salt thereof, togetherwith one or more pharmaceutically acceptable carriers or excipients.

The compounds according to the invention and their pharmaceuticallyacceptable salts have valuable pharmacological properties, particularlyan inhibitory effect on β-catenin. In view of their ability to inhibitsignaling in the Wnt pathway, and in particular to reduce the levels ofnuclear β-catenin, the compounds according to the invention and theirpharmaceutically acceptable salts are suitable for the treatment and/orprevention of any condition or disease which may be affected byover-activation of signaling in the Wnt pathway, in particular thoseconditions or diseases which involve activation of β-catenin.

The term “Wnt signaling pathway” is used to refer to the chain of eventsnormally mediated by Wnt, LRP (LDL-receptor related protein), Frizzledand β-catenin, among others, and resulting in changes in gene expressionand other phenotypic changes typical of Wnt activity.

The Wnt pathway plays a central role in the pathology of a variety ofcancers. The compounds of the invention are thus particularly suitablefor preventing and/or retarding proliferation and metastasis of tumorcells, in particular carcinomas such as adenocarcinomas. Morespecifically, the compounds are effective in treatment and/or preventionof the following cancers: colon cancers (such as colorectal cancer),pancreatic cancer (e.g. pancreas adenocarcinoma), gastric cancer, livercancers (e.g. hepatocellular and hepatoblastoma carcinomas), Wilms tumorof the kidney, medulloblastoma, skin cancers (e.g. melanoma), non-smallcell lung cancer, cervical cancer, ovarian cancers (e.g. ovarianendometrial cancer), bladder cancer, thyroid cancers (e.g. anaplasticthyroid cancer), head and neck cancer, breast cancer, prostate cancerand glioblastoma. Particularly preferably, the compounds hereindescribed may be used in the treatment and/or prevention of breastcancer, non-small cell lung cancer, ovarian, thyroid, colorectal,pancreatic and prostate cancers and glioblastoma. Treatment orprevention of breast, non-small cell lung, pancreatic and colorectalcancers represents a particularly preferred aspect of the invention.

As used herein, the term “proliferation” refers to cells undergoingmitosis. The term “retarding proliferation” indicates that the compoundsinhibit proliferation of a cancer cell. In preferred embodiments,“retarding proliferation” indicates that DNA replication is at least 10%less than that observed in untreated cells, more preferably at least 25%less, yet more preferably at least 50% less, e.g. 75%, 90% or 95% lessthan that observed in untreated cancer cells.

The term “carcinoma” refers to any malignant growth which arises fromepithelial cells. Exemplary carcinomas include basal cell carcinoma,squamous cell carcinoma and adenocarcinoma. Adenocarcinomas aremalignant tumors originating in the glandular epithelium and includecolorectal, pancreatic, breast and prostate cancers.

Viewed from a further aspect the invention thus provides a compound offormula I, II, Ia, IIa, Ib, IIb, IIc or IId, or a pharmaceuticallyacceptable salt thereof, for use in therapy.

Unless otherwise specified, the term “therapy” as used herein isintended to include both treatment and prevention.

In a still further aspect the invention provides a compound of formulaI, II, Ia, IIa, Ib, IIb, IIc or IId, or a pharmaceutically acceptablesalt thereof, for use in the treatment or prevention of colon cancers(such as colorectal cancer), pancreatic cancer, gastric cancer, livercancers (e.g. hepatocellular and hepatoblastoma carcinomas), Wilms tumorof the kidney, medulloblastoma, skin cancers (e.g. melanoma), non-smallcell lung cancer, cervical cancer, ovarian endometrial cancer, bladdercancer, anaplastic thyroid cancer, head and neck cancer, breast cancer,prostate cancer or glioblastoma.

In another aspect the invention provides the use of a compound offormula I, II, Ia, IIa, Ib, IIb, IIc or IId or a pharmaceuticallyacceptable salt thereof, in the manufacture of a medicament for use in amethod of treatment or prevention of colon cancers (such as colorectalcancer), pancreatic cancer, gastric cancer, liver cancers (e.g.hepatocellular and hepatoblastoma carcinomas), Wilms tumor of thekidney, medulloblastoma, skin cancers (e.g. melanoma), non-small celllung cancer, cervical cancer, ovarian endometrial cancer, bladdercancer, anaplastic thyroid cancer, head and neck cancer, breast cancer,prostate cancer or glioblastoma.

Also provided is a method of treatment of a human or non-human animalbody to combat or prevent colon cancers (such as colorectal cancer),pancreatic cancer, gastric cancer, liver cancers (e.g. hepatocellularand hepatoblastoma carcinomas), Wilms tumor of the kidney,medulloblastoma, skin cancers (e.g. melanoma), non-small cell lungcancer, cervical cancer, ovarian endometrial cancer, bladder cancer,anaplastic thyroid cancer, head and neck cancer, breast cancer, prostatecancer or glioblastoma, said method comprising the step of administeringto said body an effective amount of a compound of formula I, II, Ia,IIa, Ib, IIb, IIc or IId as herein defined or a pharmaceuticallyacceptable salt thereof.

Small molecules that selectively target the developmental pathways whichcontrol pattern formation during embryogenesis, including Wnt signallingpathways, are considered to be valuable for directing differentiation ofpluripotent stem cells toward many desired tissue types (see Wang etal., ACS Chemical Biology, 16 Nov. 2010). As modulators of Wntsignalling, the compounds herein described also have effects on thedevelopment of cellular differentiation. The compounds described hereintherefore have valuable properties for use in regenerative medicine, forexample in protocols for lineage specific in vitro differentiation ofprogenitor cells. By “progenitor cell” is meant a cell with the capacityto differentiate into another cell type, e.g. a stem cell.

According to this aspect, the present invention provides a method (e.g.an in vitro method) of promoting and/or directing cellulardifferentiation comprising contacting a progenitor cell with aneffective amount of a compound of formula I, II, Ia, IIa, Ib, IIb, IIcor IId as herein defined or a pharmaceutically acceptable salt thereof.In particular, the progenitor cell is contacted with said at least onecompound under suitable conditions and for a sufficient time for theprogenitor cell to differentiate into a new cell type. In a relatedaspect, the present invention provides the use of at least one compoundas herein defined for promoting and/or directing cellulardifferentiation of a progenitor cell, especially in vitro.

Preferably, the progenitor cell is a totipotent or a pluripotent cell,especially a stem cell such as an embryonic stem cell. Preferred aremammalian progenitor cells such as mouse, rat and human cells,especially human cells. Such stem cells may be obtained from establishedcell cultures or may be derived directly from mammalian tissue bymethods known in the art, including non tissue-destructive methods.

In a preferred embodiment, the progenitor cell is promoted and/ordirected to differentiate into a new cell type which is a myocyte (e.g.a cardiomyocyte), a neuronal cell (e.g. a dopaminergic neuronal cell),an endocrine pancreatic cell or a hepatocyte or a cell type which mayfurther differentiate into a myocyte, a neuronal cell, an endocrinepancreatic cell or a hepatocyte. Especially preferably, the progenitorcell is an embryonic stem cell and the new cell type is a cardiomyocyte,a dopaminergic neuronal cell, an endocrine pancreatic cell or ahepatocyte, especially a cardiomyocyte.

The dosage required to achieve the desired activity of the compoundsherein described will depend on the compound which is to beadministered, the patient, the nature and severity of the condition, themethod and frequency of administration and may be varied or adjustedaccording to choice. Typically, the dosage may be expected to be in therange from 1 to 100 mg, preferably 1 to 30 mg (when administeredintravenously) and from 1 to 1000 mg, preferably from 1 to 200 mg (whenadministered orally).

The compounds of the invention may be formulated with one or moreconventional carriers and/or excipients according to techniques wellknown in the art. Typically, the compositions will be adapted for oralor parenteral administration, for example by intradermal, subcutaneous,intraperitoneal or intravenous injection. Suitable pharmaceutical formsthus include plain or coated tablets, capsules, suspensions andsolutions containing the active component optionally together with oneor more conventional inert carriers and/or diluents, such as cornstarch, lactose, sucrose, microcrystalline cellulose, magnesiumstearate, polyvinylpyrrolidone, citric acid, tartaric acid, water,water/ethanol, water/glycerol, water/sorbitol, water/polyethyleneglycol,propylene glycol, stearylalcohol, carboxymethylcellulose or fattysubstances such as hard fat or suitable mixtures of any of the above.

The pharmacological properties of the compounds of the invention can beanalysed using standard assays for functional activity. Detailedprotocols for testing of the compounds of the invention are provided inthe Examples.

The invention will now be described in more detail in the followingnon-limiting Examples and Figures, in which:

FIG. 1 shows the effect of a compound of the invention on cell growth inRKO, HCT-15, WiDr, HT29, DLD-1, COLO320DM and COLO205 cells (FIGS. 1A to1G, respectively);

FIG. 2 shows the effect of XAV939 (FIG. 2A) and a compound of theinvention (FIG. 2B) on the inhibition of TNKS1 in vitro;

FIG. 3 shows the effect of XAV939 (FIG. 3A) and a compound of theinvention (FIG. 3B) on the inhibition of TNKS2 in vitro;

FIG. 4 shows the effect of XAV939 (FIG. 4A) and a compound of theinvention (FIG. 4B) on the inhibition of PARP in vitro; and

FIG. 5 shows the results of immunostaining of SW480 cells treated withDMSO (negative control, left-hand panels) and a compound of theinvention (right-hand panels).

Examples 6 to 9 are examples of formulations in which reference to the“active substance” denotes one or more compounds according to theinvention, including the salts thereof.

EXAMPLE 1 Preparation of4-(5-((E)-2-(5-(2,4-dichlorophenyl)-4-(2-chlorophenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (1))

(a) Preparation of 2,4-dichloro-N-(2-chlorophenyl)benzamide 2

To a solution of 2-chloroaniline (5.15 g, 40.37 mmol) in dichloromethane(50 mL) was added 2,4-dichlorobenzoyl chloride 1 (8.88 g, 42.39 mmol)and then triethylamine (6.1 mL, 44 mmol). The reaction mixture wasstirred overnight at ambient temperature and ethyl acetate (150 mL) wasadded followed by 1N HCl (50 mL). The organic phase was washed withwater and brine, dried, and concentrated under reduced pressure to yielda light yellow solid. 2,4-dichloro-N-(2-chlorophenyl)benzamide 2 wasused for the next step without further purification. Yield: 11 g, 90%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.52 (dd, 2H), 7.62 (d, 1H), 7.52 (s,1H), 7.38-7.22 (m, 3H), 7.08 (d, 1H).

(b) Preparation of N′-(2-chlorophenyl)-4-fluorobenzimidohydrazide 4

A mixture of 2,4-dichloro-N-(2-chlorophenyl)benzamide 2 (6.0 g, 20 mmol)and PCl₅ (4.58 g, 22 mmol) in benzene (40 mL) was stirred under refluxfor 16 hours. The solvent was removed and further dried under highvaccum. The residue2-chloro-N-(chloro(2,4-dichlorophenyl)methylene)benzenamine 3 was cooleddown by ice water bath. Anhydrous THF was added (20 mL) followed byhydrazine monohydrate (5.0 mL) and the reaction mixture was furtherstirred at ambient temperature for 3 hours. After the solvent wasremoved, the residue was mixed with ether:hexane (1:1, 30 mL), the solidwas filtered and further washed with ether:hexane mixture to yield apure solid as N-amino-2,4-dichloro-N′-(2-chlorophenyl)benzamidine 4.Yield: 4.4 g, 70%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 7.43 (d, 1H), 7.38-7.22 (m, 3H), 6.98(dd, 1H), 6.82 (dd, 1H), 6.41 (d, 1H).

(c) Preparation of 2-methoxyisonicotinohydrazide 6

To a solution of 2-methoxypyridine-4-carboxylic acid 5 (1.0 g, 6.5 mmol)in CH₃OH (30 mL) was added a few drops of SOCl₂ at ambient temperature.The mixture was stirred under reflux for 16 hours and concentrated togive the ester. To the solution of the ester in CH₃OH was addedhydrazine monohydrate (2.0 mL, 37 mmol) at ambient temperature. Thereaction mixture was stirred at ambient temperature for 16 hours andconcentrated to give 2-methoxyisonicotinohydrazide 6 as a white solid.Yield: 0.87 g, 80%.

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm): 10.0 (bs, 1H), 8.25 (dd, 1H), 7.31(dd, 1H), 7.13 (s, 1H), 4.66 (bs, 2H), 3.86 (s, 3H).

(d) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 8

To a solution of 2-methoxyisonicotinohydrazide 6 (0.80 g, 4.89 mmol) inTHF (20 mL) was added malenic anhydride 7 (0.47 g, 4.79 mmol). Themixture was stirred at ambient temperature for 16 hours. The solid wasfiltered and dried under vacuum to afford the pure compound(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 8 as awhite solid. Yield: 1.02 g, 80%.

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm): 8.32 (d, 1H), 7.38 (d, 1H), 7.22 (s,1H), 6.38 (dd, 2H), 3.90 (s, 3H), 3.40 (bs, 3H).

(e) Preparation of4-(5-((E)-2-(5-(2,4-dichlorophenyl)-4-(2-chlorophenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (1))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 8 (0.2 g,0.75 mmol) in CH₃CN was added POCl₃ (0.37 g, 2.4 mmol). The mixture wasstirred under reflex for 16 hours, cooled to ambient temperature andconcentrated under reduced pressure. The residue was dissolved in CH₃CN,treated with a solution ofN-amino-2,4-dichloro-N′-(2-chlorophenyl)benzamidine 4 (0.38 g, 1.2 mmol)in CH₃CN and K₂CO₃ (0.38 g, 1.2 mmol). The mixture was stirred atambient temperature for 1 hour, refluxed for 2 hours and filtered. Thefiltrate was concentrated. The residue was subjected to flash columnchromatography (FCC) (50-60% EtOAc/hexanes) to afford the title compound4-(5-((E)-2-(5-(2,4-dichlorophenyl)-4-(2-chlorophenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridineas a white solid. Yield: 0.11 g, 28%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.38 (d, 1H), 7.62-7.18 (m, 11H), 4.03(s, 3H).

MS (M+1) 525.3.

HPLC (Waters 625 LC System): 95%.

EXAMPLE 2 Preparation of4-(5-((E)-2-(4-(2-chlorophenyl)-5-(4-(methylsulfonyl)phenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (2))

(a) Preparation of N-(2-chlorophenyl)-4-(methylsulfonyl)benzamide 3

To a solution of 4-(methylsulfonyl)benzoic acid 1 (1.0 g, 5 mmol) in 50mL of dichloromethane was added thionyl chloride (5 mL). The mixture wasstirred at ambient temperature for 18 hours. After removal of allsolvent, the residue was dissolved in 50 mL of dichloromethane. To thesolution was added o-chloroaniline (0.766 g, 6 mmol) followed by 1.4 mLof Et₃N. The mixture was stirred at ambient temperature for 2 hours.After washing with 1N HCl and water, the organic layer was concentratedto give N-(2-chlorophenyl)-4-(methylsulfonyl)benzamide 3 as a solid.Yield: 1.18 g, 76%.

¹HNMR (CDCl₃) δ (ppm): 8.54 (d, 1H); 8.43 (bs, 1H); 8.12 (m, 4H); 7.46(d, 1H); 7.38 (dd, 1H); 7.15 (dd, 1H); 3.10 (s, 3H).

(b) Preparation ofN-amino-N′-(2-chlorophenyl)-4-methylsulfonyl-benzamidine 5

To a suspension of N-(2-chlorophenyl)-4-(methylsulfonyl)benzamide 3 (1.0g, 3.23 mmol) in 50 mL of C₆H₆ was added 10 mL of POCl₃. The mixture wasrefluxed for 15 hours. After removal of solvent, the residue wasdissolved in 50 mL of THF and 3 mL of H₂NNH₂.H₂O added. The mixture wasstirred at ambient temperature for 2 hours. After removal of solvent,the residue was purified by column (2-5% methanol in dichloromethane) togive N-amino-N′-(2-chlorophenyl)-4-methylsulfonyl-benzamidine 5 as asolid. Yield: 0.8 g, 77%.

¹HNMR (CDCl₃) δ (ppm): 7.90 (d, 2H); 7.75 (d, 2H); 7.40 (d, 1H); 7.05(dd, 1H); 6.85 (dd, 1H); 6.40 (d, 1H); 5.95 (br, 1H); 5.70 (br, 2H);3.3.05 (s, 3H).

(c) Preparation of 2-methoxypyridine-4-carbohydrazide 8

To a solution of 2-methoxypyridine-4-carboxylic acid 6 (2.0 g, 13 mmol)in 30 mL of MeOH was added 3 mL of thionyl chloride. The mixture wasstirred under reflux for 2 hours. After removal of the solvent, theresidue was dissolved in 30 mL of THF and 3 mL of H₂NNH₂.H₂O added. Themixture was heated at reflux for 0.5 hours. After removal of solvent,the residue was purified by column (5-10% of methanol indichloromethane) to give 2-methoxypyridine-4-carbohydrazide 8 as asolid. Yield: 1.76 g, 81%.

¹HNMR (DMSO-d₆) δ (ppm): 9.98 (s, 1H); 8.25 (d, 1H); 7.30 (d, 1H); 7.13(s, 1H); 4.57 (s, 2H); 3.86 (s, 3H).

(d) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9

To a solution of compound 8 (167 mg, 1.0 mmol) in 10 mL of THF was addedfuran-2,5-dione (110 mg, 1.1 mmol). The mixture was stirred at ambienttemperature for 2 hours. After removal of the solvent, the residue wasdiluted with 10 mL of dichloromethane. The solid was collected byfiltration to give(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 as asolid.

Yield: 190 mg, 71%.

¹HNMR (DMSO-d₆) δ (ppm): 10.45 (s, 2H); 8.32 (d, 1H); 7.36 (d, 1H); 7.20(s, 1H); 6.41 (d, 1H); 6.29 (d, 1H); 3.90 (s, 3H).

(e) Preparation of4-(5-((E)-2-(4-(2-chlorophenyl)-5-(4-(methylsulfonyl)phenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (2))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 (180 mg,0.68 mmol) in 20 mL of PhMe was added 0.2 mL of POCl₃. The mixture wasstirred at reflux for 2 hours. After removal of solvent, the residue wasdissolved in 20 mL of THF and 220 mg (0.68 mmol) ofN-amino-N′-(2-chlorophenyl)-4-methylsulfonyl-benzamidine 5 added. Themixture was stirred at ambient temperature for 18 hours. After removalof solvent, the residue was purified by column (0-5% of methanol indichloromethane) to give4-(5-((E)-2-(4-(2-chlorophenyl)-5-(4-(methylsulfonyl)phenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridineas a solid. Yield: 30 mg, 8.3%.

¹HNMR (CDCl₃) δ (ppm): 8.32 (d, 1H); 7.90 (d, 2H); 7.80-7.40 (m, 8H);7.32 (m, 1H); 7.10 (d, 1H); 4.00 (s, 3H); 3.3.05 (s, 3H).

MS (M+1) 535.1.

HPLC (Waters 625 LC System): 96%.

EXAMPLE 3 Preparation of4-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyridine(Compound (3))

(a) Preparation of 2-methoxypyridine-4-carbohydrazide 3

To a solution of 2-methoxypyridine-4-carboxylic acid 1 (1.0 g, 6.5 mmol)in CH₃OH (30 mL) was added a few drops of SOCl₂ at ambient temperature.The mixture was stirred under reflux for 16 hours and concentrated togive methyl 2-methoxypyridine-4-carboxylate 2. To the solution of2-methoxypyridine-4-carboxylate 2 in CH₃OH was added hydrazinemonohydrate (2.0 mL, 37 mmol) at ambient temperature. The reactionmixture was stirred at ambient temperature for 16 hours and concentratedto give 2-methoxypyridine-4-carbohydrazide 3 as a white solid. Yield:0.87 g, 80%.

¹H-NMR (300 MHZ, DMSO-d₆) δ (ppm): 10.0 (bs, 1H), 8.25 (dd, 1H), 7.31(dd, 1H), 7.13 (s, 1H), 4.66 (bs, 2H), 3.86 (s, 3H).

(b) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4

To a solution of 2-methoxypyridine-4-carbohydrazide 3 (0.80 g, 4.89mmol) in THF (20 mL) was added malenic anhydride 7 (0.47 g, 4.79 mmol).The mixture was stirred at ambient temperature for 16 hours. The solidwas filtered and dried under vacuum to afford pure compound(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 as awhite solid. Yield: 1.02 g, 80%.

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm): 8.32 (d, 1H), 7.38 (d, 1H), 7.22 (s,1H), 6.38 (dd, 2H), 3.90 (s, 3H), 3.40 (bs, 3H).

(c) Preparation of N-(2-chlorophenyl)isonicotinamide 6

A mixture of isonicotinic acid 5 (1.0 g, 8.12 mmol) and SOCl₂ (2.4 mL)in DCM (10 mL) was stirred for 2 hours and concentrated. The residue wasdissolved in DCM (20 mL). To the solution was added 2-chlorobenzenamine(1.7 g, 8.15 mmol) and then triethylamine (1.4 mL, 10 mmol). Thereaction mixture was stirred overnight at ambient temperature and ethylacetate (50 mL) was added followed by 1N HCl (10 mL). The organic phasewas washed with water and brine, dried and concentrated under reducedpressure to yield a light yellow solid. Yield: 1.5 g, 80%.N-(2-chlorophenyl)isonicotinamide 6 was used for the next step withoutfurther purification.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.84 (dd, 2H), 8.51 (d, 1H), 8.48 (d,1H), 7.75 (dd, 2H), 7.43 (dd, 1H), 7.38-7.32 (m, 1H), 7.16-7.10 (m, 1H).

(d) Preparation of N-amino-N′-(2-chlorophenyl)pyridine-4-carboxamidine 8

A mixture of N-(2-chlorophenyl)isonicotinamide 6 (1.5 g, 6.45 mmol) andPCl₅ (1-6 g, 7.8 mmol) in benzene (40 mL) was stirred under reflux for16 hours. The solvent was removed and further dried under high vaccum.The residue (Z)-2-chloro-N-(chloro(pyridin-4-yl)methylene)benzenamine 7was cooled down by ice water bath and anhydrous THF added (20 mL)followed hydrazine monohydrate (1.2 mL). The reaction mixture wasfurther stirred at ambient temperature for 3 hours. After the solventwas removed the residue was mixed with ether:hexane (1:1, 30 mL), thesolid was filtered and further washed with ether:hexane mixture to yielda pure solid as N-amino-N′-(2-chlorophenyl)pyridine-4-carboxamidine 8.Yield: 1.1 g, 70%.

¹H-NMR (300 Hz, CDCl₃) δ 8.55 (dd, 2H), 7.43 (dd, 2H), 7.38 (d, 1H),7.07-7.01 (m, 1H), 6.86-6.80 (dd, 1H), 6.38 (dd, 1H), 5.66 (bs, 3H).

(e) Preparation of4-(5-((E)-2-(5-(2,4-dichlorophenyl)-4-(2-chlorophenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (3))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 (0.2 g,0.75 mmol) in CH₃CN was added POCl₃ (0.37 g, 2.4 mmol). The mixture wasstirred under reflux for 16 hours, cooled to ambient temperature andconcentrated under reduced pressure. The residue was dissolved in CH₃CNand treated with a solution ofN-amino-N′-(2-chlorophenyl)pyridine-4-carboxamidine 8 (0.38 g, 1.5 mmol)in CH₃CN and K₂CO₃ (0.38 g, 1.2 mmol). The mixture was stirred atambient temperature for 1 hour, refluxed for 2 hours and filtered. Thefiltrate was concentrated. The residue was subjected to FCC (50-60%EtOAc/hexanes) to afford the title compound4-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyridineas a white solid. Yield: 0.11 g, 28%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.58 (d, 2H), 8.32 (d, 1H), 7.74-7.64(m, 3H), 7.60-7.55 (m, 1H), 7.50-7.43 (m, 2H), 7.35-7.31 (m, 3H), 7.09(d, 1H), 3.98 (s, 3H).

MS (M+1) 457.4644.

HPLC (Waters 625 LC System): 96%.

EXAMPLE 4 Preparation of5-chloro-2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyridine(Compound (4))

(a) Preparation of 2-methoxypyridine-4-carbohydrazide 3

To a solution of 2-methoxypyridine-4-carboxylic acid 1 (1.0 g, 6.5 mmol)in CH₃OH (30 mL) was added a few drops of SOCl₂ at ambient temperature.The mixture was stirred under reflux for 16 hours and concentrated togive methyl 2-methoxypyridine-4-carboxylate 2. To the solution of2-methoxypyridine-4-carboxylate 2 in CH₃OH was added hydrazinemonohydrate (2.0 mL, 37 mmol) at ambient temperature. The reactionmixture was stirred at ambient temperature for 16 hours and concentratedto give 2-methoxypyridine-4-carbohydrazide 3 as a white solid. Yield:0.87 g, 80%.

¹H-NMR (300 MHZ, DMSO-d₆) δ (ppm): 10.0 (bs, 1H), 8.25 (dd, 1H), 7.31(dd, 1H), 7.13 (s, 1H), 4.66 (bs, 2H), 3.86 (s, 3H).

(b) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4

To a solution of 2-methoxypyridine-4-carbohydrazide 3 (0.80 g, 4.89mmol) in THF (20 mL) was added malenic anhydride 7 (0.47 g, 4.79 mmol).The mixture was stirred at ambient temperature for 16 hours. The solidwas filtered and dried under vacuum to afford pure compound(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 as awhite solid. Yield: 1.02 g, 80%.

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm): 8.32 (d, 1H), 7.38 (d, 1H), 7.22 (s,1H), 6.38 (dd, 2H), 3.90 (s, 3H), 3.40 (bs, 3H).

(b) Preparation of 5-chloro-N-(2-chlorophenyl)pyridine-2-carboxamide 6

A mixture of 5-chloropyridine-2-carboxylic acid 5 (1.0 g, 6.35 mmol) andSOCl₂ (2.4 mL) in DCM (10 mL) was stirred for 2 hours and concentrated.The residue was dissolved in DCM (20 mL). To the solution was added2-chlorobenzenamine (1.0 g, 7.84 mmol) and then triethylamine (2.0 mL,14.43 mmol). The reaction mixture was stirred overnight at ambienttemperature and ethyl acetate (50 mL) was added followed by 1N HCl (10mL). The organic phase was washed with water and brine, dried andconcentrated under reduced pressure to yield a light yellow solid.Yield: 0.8 g, 47%. 5-chloro-N-(2-chlorophenyl)pyridine-2-carboxamide 6was used for the next step without further purification.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.5 (bs, 1H), 8.65-8.60 (m, 2H), 8.28(d, 1H), 7.90 (dd, 1H), 7.44 (dd, 1H), 7.38-7.30 (m, 1H), 7.16-7.08 (m,1H).

(c) Preparation ofN-amino-5-chloro-N′-(2-chlorophenyl)pyridine-2-carboxamidine 8

A mixture of 5-chloro-N-(2-chlorophenyl)pyridine-2-carboxamide 6 (0.8 g,3.0 mmol) and PCl₅ (1 g, 4.8 mmol) in benzene (30 mL) was stirred underreflux for 16 hours. The solvent was removed and further dried underhigh vaccum. The residue(2Z)-5-chloro-N-(2-chlorophenyl)pyridine-2-carboximidoyl chloride 7 wascooled down by ice water bath and anhydrous THF added (20 mL) followedby hydrazine monohydrate (1.0 mL). The reaction mixture was furtherstirred at ambient temperature for 3 hours. After the solvent wasremoved the residue was mixed with ether:hexane (1:1, 30 mL), the solidwas filtered and further washed with ether:hexane mixture to yield apure solid asN-amino-5-chloro-N′-(2-chlorophenyl)pyridine-2-carboxamidine 8. Yield:0.5 g, 60%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.43 (dd, 1H), 7.98 (d, 1H), 7.65 (dd,1H), 7.41-7.32 (m, 2H), 7.19-7.13 (m, 1H), 6.90-6.85 (m, 1H), 6.50 (dd,1H), 5.36 (bs, 2H).

(d) Preparation of5-chloro-2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyridine(Compound (4))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 (0.2 g,0.75 mmol) in CH₃CN was added POCl₃ (0.37 g, 2.4 mmol). The mixture wasstirred under reflux for 16 hours, cooled to ambient temperature andconcentrated under reduced pressure. The residue was dissolved in CH₃CN,treated with a solution ofN-amino-5-chloro-N′-(2-chlorophenyl)pyridine-2-carboxamidine 8 (0.38 g,1.2 mmol) in CH₃CN and K₂CO₃ (0.38 g, 1.2 mmol). The mixture was stirredat ambient temperature for 1 hour, refluxed for 2 hours, and filtered.The filtrate was concentrated. The residue was subjected to FCC (50-60%EtOAc/hexanes) to afford the title compound5-chloro-2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyridineas a white solid. Yield: 0.11 g, 28%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.33 (dd, 2H), 8.18 (d, 1H), 7.78 (dd,1H), 7.60-7.39 (m, 6H), 7.31 (dd, 1H), 7.09 (d, 1H), 3.98 (s, 3H).

MS (M+1) 491.4821.

HPLC (Waters 625 LC System): 94%.

EXAMPLE 5 Preparation of4-(5-((E)-2-(4-(2-chlorophenyl)-5-(3-methyl-1,2,4-oxadiazol-5-yl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (5))

(a) Preparation of 2-methoxypyridine-4-carbohydrazide 3

To a solution of 2-methoxypyridine-4-carboxylic acid 1 (1.0 g, 6.5 mmol)in CH₃OH (30 mL) was added a few drops of SOCl₂ at ambient temperature.The mixture was stirred under reflux for 16 hours and concentrated togive methyl 2-methoxypyridine-4-carboxylate 2. To the solution of2-methoxypyridine-4-carboxylate 2 in CH₃OH was added hydrazinemonohydrate (2.0 mL, 37 mmol) at ambient temperature. The reactionmixture was stirred at ambient temperature for 16 hours and concentratedto give 2-methoxypyridine-4-carbohydrazide 3 as a white solid. Yield:0.87 g, 80%.

¹H-NMR (300 MHZ, DMSO-d₆) δ (ppm): 10.0 (bs, 1H), 8.25 (dd, 1H), 7.31(dd, 1H), 7.13 (s, 1H), 4.66 (bs, 2H), 3.86 (s, 3H).

(b) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4

To a solution of 2-methoxypyridine-4-carbohydrazide 3 (0.80 g, 4.89mmol) in THF (20 mL) was added malenic anhydride 7 (0.47 g, 4.79 mmol).The mixture was stirred at ambient temperature for 16 hours. The solidwas filtered and dried under vacuum to afford pure compound(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 as awhite solid. Yield: 1.02 g, 80%.

¹H-NMR (300 MHz, DMSO-d₆) δ (ppm): 8.32 (d, 1H), 7.38 (d, 1H), 7.22 (s,1H), 6.38 (dd, 2H), 3.90 (s, 3H), 3.40 (bs, 3H).

(c) Preparation ofN-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide 6

To a mixture of ethyl 3-methyl-1,2,4-oxadiazole-5-carboxylate 5 (1.0 g,6.4 mmol) and 2-chlorobenzenamine (1.23 g, 9.6 mmol) in toluene (20 mL)was added AlMe₃ (2.0 M 4.8 mL, 9.6 mmol) dropwise. The reaction mixturewas stirred under reflux overnight, treated with 1N HCl (5 mL) and ethylacetate (50 mL). The organic phase was dried and concentrated. Theresidue was purified by column chromatography eluting with hexane:ethylacetate (1:3) to affordN-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide 6. Yield: 1.0g, 66%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.98 (s, 0.5H), 9.38 (s, 0.5H),8.52-8.43 (m, 1H), 7.48-7.38 (dd, 1H), 7.40-7.32 (m, 1H), 7.22-7.13 (m,1H), 2.56 (s, 3H).

(d) Preparation ofN-amino-N′-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamidine 8

A mixture of N-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide6 (1.0 g, 4.21 mmol) and PCl₅ (2 g, 9.6 mmol) in benzene (40 mL) wasstirred under reflux for 16 hours. The solvent was removed and furtherdried under high vaccum. The residue(Z)-2-chloro-N-(chloro(3-methyl-1,2,4-oxadiazol-5-yl)methylene)benzenamine7 was cooled down by ice water bath and anhydrous THF added (20 mL)followed hydrazine monohydrate (5.0 mL). The reaction mixture wasfurther stirred at ambient temperature for 3 hours. After the solventwas removed the residue was mixed with ether:hexane (1:1, 30 mL), thesolid was filtered and further washed with ether:hexane mixture to yielda pure solid asN-amino-N′-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamidine 8.Yield: 0.5 g, 47%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 7.40 (t, 1H), 7.20 (t, 1H), 6.92-6.82(m, 1H), 6.76 (s, 0.5H), 6.52-6.43 (m, 1H), 6.28 (s, 0.5H), 6.12 (s,1H), 5.25 (s, 1H), 2.43 (s, 3H).

(e) Preparation of4-(5-((E)-2-(5-(2,4-dichlorophenyl)-4-(2-chlorophenyl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridine(Compound (5))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 4 (0.2 g,0.75 mmol) in CH₃CN was added POCl₃ (0.37 g, 2.4 mmol). The mixture wasstirred under reflux for 16 hours, cooled to ambient temperature andconcentrated under reduced pressure. The residue was dissolved in CH₃CN,treated with a solution ofN-amino-N′-(2-chlorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamidine 8(0.5 g, 1.99 mmol) in CH₃CN and K₂CO₃ (0.38 g, 1.2 mmol). The mixturewas stirred at ambient temperature for 1 hour, refluxed for 2 hours, andfiltered. The filtrate was concentrated. The residue was subjected toFCC (50-60% EtOAc/hexanes) to afford the title compound4-(5-((E)-2-(4-(2-chlorophenyl)-5-(3-methyl-1,2,4-oxadiazol-5-yl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)-2-methoxypyridineas a white solid. Yield: 0.1 g, 28%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.32 (d, 1H), 7.75-7.69 (m, 2H),7.60-7.55 (m, 1H), 7.50-7.45 (m, 1H), 7.30 (s, 1H), 7.08 (d, 1H), 3.98(s, 3H), 2.35 (s, 3H).

MS (M+1) 462.5578.

HPLC (Waters 625 LC System): 98%.

EXAMPLE 6 Tablets Containing 100 mg of Active Substance

Each tablet contains:

active substance 100.0 mg lactose 80.0 mg corn starch 34.0 mgpolyvinylpyrrolidone 4.0 mg magnesium stearate 2.0 mg

The active substance, lactose and corn starch are mixed together anduniformly moistened with an aqueous solution of thepolyvinylpyrrolidone. The moist composition is screened (2.0 mm meshsize) and dried at 50° C. The lubricant is added and the final mixtureis compressed to form tablets. Final weight of each tablet is 220 mg.

EXAMPLE 7 Tablets Containing 150 mg of Active Substance

Each tablet contains:

active substance 150.0 mg powdered lactose 89.0 mg corn starch 40.0 mgcolloidal silica 10.0 mg polyvinylpyrrolidone 10.0 mg magnesium stearate1.0 mg

The active substance is mixed with lactose, corn starch and silica andmoistened with an aqueous polyvinylpyrrolidone solution. The moistcomposition is passed through a screen with a mesh size of 1.5 mm. Theresulting granules are dried at 45° C., then mixed with the magnesiumstearate. Tablets are pressed from the mixture. Each tablet weighs 300mg.

EXAMPLE 8 Ampoules Containing 10 mg Active Substance

Each ampoule contains:

active substance 10.0 mg 0.01N hydrochloric acid q.s. double-distilledwater ad 2.0 ml

The active substance is dissolved in the necessary amount of 0.01 N HCl,made isotonic with common salt, sterile filtered and transferred into 2ml ampoules.

EXAMPLE 9 Ampoules Containing 50 mg of Active Substance

Each ampoule contains:

active substance 50.0 mg 0.01N hydrochloric acid q.s. double-distilledwater ad 10.0 ml

The active substance is dissolved in the necessary amount of 0.01 N HCl,made isotonic with common salt, sterile filtered and transferred into 10ml ampoules.

EXAMPLE 10 Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-ethoxypyridine(Compound (13))

(a) Preparation of ethyl 5-bromopyridine-2-carboxylate 2

To a suspension of 5-bromopyridine-2-carboxylic acid 1 (2.02 g, 10 mmol)in 50 mL of EtOH was added thionyl chloride (5 mL). The mixture wasstirred at ambient temperature for 18 h. After removal of all solvent,the residue was purified by column (2:1 of hexane/ethyl acetate) to give2.3 g of ethyl 5-bromopyridine-2-carboxylate 2 as an oil. Yield: 2.3 g,quantitative yield. ¹HNMR (CDCl₃) δ (ppm): 8.80 (d, 1H), 8.00 (m, 2H),4.50 (m, 2H), 1.44 (t, 3H).

(b) Preparation of ethyl 5-ethoxypyridine-2-carboxylate 3

To a solution of ethyl 5-bromopyridine-2-carboxylate 2 (1.5 g, 6.5 mmol)in 20 mL of EtOH was added a solution of sodium (0.18 g, 7.8 mmol) in 20mL of EtOH. The mixture was stirred at reflux for 3 h. After removal ofall solvent, the residue was purified by column (2:1 of hexane/ethylacetate) to give ethyl 5-ethoxypyridine-2-carboxylate 3 as an oil.Yield: 0.26 g, 20%. ¹HNMR (CDCl₃) δ (ppm): 8.38 (d, 1H), 8.10 (d, 1H),8.23 (d, 1H), 7.22 (dd, 1H), 4.45 (m, 2H), 4.25 (m, 1H), 1.40 (m, 6H).

(c) Preparation of N-(2-chlorophenyl)-5-ethoxypyridine-2-carboxamide 4

To a solution of o-chloroaniline (0.68 g, 5.3 mmol) in 20 mL of PhMe wasadded 3 mL of AlMe₃. The mixture was stirred at ambient temperature for0.5 h and added into a solution of ethyl 5-ethoxypyridine-2-carboxylate3 (0.26 g, 1.33 mmol) in 10 mL of PhMe. The mixture was stirred atambient temperature for 15 h and quenched with 20 mL of 1N HCl. Themixture was extracted with ethyl acetate. The organic layer wasseparated and concentrated. The residue was purified by column (2:1 ofhexane/ethyl acetate) to giveN-(2-chlorophenyl)-5-ethoxypyridine-2-carboxamide 4 as a solid. Yield:310 mg, 96%.

¹HNMR (CDCl₃) δ (ppm): 10.55 (s, 1H), 8.65 (d, 1H), 8.30 (d, 1H), 8.23(d, 1H), 7.42 (d, 1H), 7.30 (m, 2H), 7.05 (dd, 1H), 4.20 (m, 2H), 1.50(t, 3H).

(d) Preparation ofN-amino-N-(2-chlorophenyl)-5-ethoxy-pyridine-2-carboxamidine 5

To a solution of N-(2-chlorophenyl)-5-ethoxypyridine-2-carboxamide 4(0.15 g, 0.64 mmol) in 20 mL of benzene was added 650 mg of PCl₅. Themixture was stirred at reflux for 2 h. After removal of the solvent, theresidue was dissolved in 30 mL of THF and added to 3 mL of H₂NNH₂.H₂O.The mixture was heated at reflux for 0.5 h. After removal of solvent,the residue was purified by column (5-10% of methanol indichloromethane) to giveN-amino-N′-(2-chlorophenyl)-5-ethoxy-pyridine-2-carboxamidine 5 as asolid. Yield: 140 mg, 95%. ¹HNMR (CDCl₃) δ (ppm): 8.20 (d, 1H), 7.95 (d,1H), 7.50 (s, 1H), 7.35 (d, 1H), 7.20 (m, 2H), 6.85 (dd, 1H), 6.55 (d,1H), 5.20 (s, 2H), 4.10 (m, 2H), 1.40 (t, 3H).

(e) Preparation of 2-methoxypyridine-4-carbohydrazide 8

To a solution of 2-methoxypyridine-4-carboxylic acid 6 (2.0 g, 13 mmol)in 30 mL of MeOH was added 3 mL of thionyl chloride. The mixture wasstirred at reflux for 2 h. After removal of the solvent, the residue wasdissolved in 30 mL of THF and added to 3 mL of H₂NNH₂.H₂O. The mixturewas heated at reflux for 0.5 h. After removal of solvent, the residuewas purified by column (5-10% of methanol in dichloromethane) to give2-methoxypyridine-4-carbohydrazide 8 as a solid. Yield: 1.76 g, 81%.

¹HNMR (DMSO-d₆) δ (ppm): 9.98 (s, 1H), 8.25 (d, 1H), 7.30 (d, 1H), 7.13(s, 1H), 4.57 (s, 2H), 3.86 (s, 3H).

(f) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9

To a solution of 2-methoxypyridine-4-carbohydrazide 8 (167 mg, 1.0 mmol)in 10 mL of THF was added maleic anhydride (110 mg, 1.1 mmol). Themixture was stirred at ambient temperature for 2 h. After removal of thesolvent, the residue was diluted with 10 mL of dichloromethane. Thesolid was collected by filtration to give(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 as asolid. Yield: 190 mg, 71%.

¹HNMR (DMSO-d₆) δ (ppm): 10.80 (s, 2H), 8.32 (d, 1H), 7.36 (d, 1H), 7.20(s, 1H), 6.41 (d, 1H), 6.29 (d, 1H), 3.90 (s, 3H).

(g) Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-ethoxypyridine(Compound (13))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 (0.30 g,mmol) in 20 mL of PhMe was added 0.5 mL of POCl₃. The mixture wasstirred at reflux for 2 h. After removal of solvent, the residue wasdissolved in 20 mL of dichloromethane and added to 240 mg (0.68 mmol) ofN-amino-N′-(2-chlorophenyl)-5-ethoxy-pyridine-2-carboxamidine 5. Themixture was stirred at ambient temperature for 2 h. After removal ofsolvent, the residue was suspended in 20 mL of PhMe and refluxed for 3h. After removal of solvent, the residue was purified by column (0-5% ofmethanol in dichloromethane) to give2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-ethoxypyridineas a solid. Yield: 20 mg, 3.5%.

¹HNMR (CDCl₃) δ (ppm): 8.33 (d, 1H), 8.24 (d, 1H), 7.89 (d, 1H),7.60-7.20 (m, 8H), 7.14 (d, 1H), 4.10 (m, 5H), 1.40 (t, 3H).

MS: 502.3 (MH⁺).

HPLC: 99%

EXAMPLE 11 Preparation of4-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyrimidine(Compound (11))

(a) Preparation of N-(2-chlorophenyl)pyrimidine-4-carboxamide 3

To a suspension of pyrimidine-4-carboxylic acid 1 (0.5 g, 4 mmol) in 30mL of dichloromethane was added thionyl chloride (3 mL). The mixture wasstirred at ambient temperature for 18 h. After removal of all solvent,the residue was suspended in 50 mL of dichloromethane and added to 1.0 g(8 mmol) of o-chloroaniline. The mixture was stirred at ambienttemperature for 16 h.

After removal of solvent, the residue was purified by column (2:1 ofhexane/ethyl acetate) to give N-(2-chlorophenyl)pyrimidine-4-carboxamide3 as a solid. Yield: 0.28 g, 30%. ¹HNMR (CDCl₃) δ (ppm): 10.60 (s, 1H);9.35 (d, 1H); 9.10 (d, 1H); 8.65 (d, 1H); 8.45 (d, 1H); 7.50-7.10 (m,3H).

(b) Preparation of N-amino-N′-(2-chlorophenyl)pyrimidine-4-carboxamidine5

To a suspension of N-(2-chlorophenyl)pyrimidine-4-carboxamide 3 (0.2 g,0.86 mmol) in 20 mL of PhMe was added 0.65 g of PCl₅. The mixture wasstirred at reflux for 3 h. To this solution was added 3 mL ofNH₂NH₂.H₂O. The mixture was stirred at ambient temperature for 2 h.After removal of all solvent, the residue was purified by column (2:1 ofhexane/ethyl acetate) to giveN-amino-N′-(2-chlorophenyl)pyrimidine-4-carboxamidine 5 as a solid.Yield: 0.15 g, 70%. ¹HNMR (DMSO-d₆) δ (ppm): 9.15 (s, 1H); 8.70 (d, 1H);7.98 (d, 1H); 7.35 (m, 1H); 7.15 (m, 1H); 6.80 (m, 1H); 6.45 (d, 1H);5.60 (s, 2H).

(c) Preparation of 2-methoxypyridine-4-carbohydrazide 8

To a solution of 2-methoxypyridine-4-carboxylic acid 6 (2.0 g, 13 mmol)in 30 mL of MeOH was added 3 mL of thionyl chloride. The mixture wasstirred at reflux for 2 h. After removal of the solvent, the residue wasdissolved in 30 mL of THF and added to 3 mL of H₂NNH₂.H₂O. The mixturewas heated at reflux for 0.5 h. After removal of solvent, the residuewas purified by column (5-10% of methanol in dichloromethane) to give2-methoxypyridine-4-carbohydrazide 8 as a solid. Yield: 1.76 g, 81%.

¹HNMR (DMSO-d₆) δ (ppm): 9.98 (s, 1H); 8.25 (d, 1H); 7.30 (d, 1H); 7.13(s, 1H); 4.57 (s, 2H); 3.86 (s, 3H).

(d) Preparation of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9

To a solution of 2-methoxypyridine-4-carbohydrazide 8 (167 mg, 1.0 mmol)in 10 mL of THF was added maleic anhydride (110 mg, 1.1 mmol). Themixture was stirred at ambient temperature for 2 h. After removal of thesolvent, the residue was diluted with 10 mL of dichloromethane. Thesolid was collected by filtration to give(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 as asolid. Yield: 190 mg, 71%.

¹HNMR (DMSO-d₆) δ (ppm): 10.45 (s, 2H); 8.32 (d, 1H); 7.36 (d, 1H); 7.20(s, 1H); 6.41 (d, 1H); 6.29 (d, 1H); 3.90 (s, 3H).

(e) Preparation of4-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyrimidine(Compound (11))

To a suspension of(Z)-4-(2-methoxypyridine-4-carboamido)-4-oxobut-2-enoic acid 9 (200 mg,0.75 mmol) in 20 mL of PhMe was added 0.5 mL of POCl₃. The mixture wasstirred at reflux for 2 h. After removal of solvent, the residue wasdissolved in 20 mL of dichloromethane and added to 150 mg (0.6 mmol) ofN-amino-N′-(2-chlorophenyl)pyrimidine-4-carboxamidine 5. The mixture wasstirred at ambient temperature for 2 h. After removal of solvent, theresidue was suspended in 20 mL of PhMe and refluxed for 3 h. Afterremoval of solvent, the residue was purified by column (0-5% of methanolin dichloromethane) to give4-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)pyrimidineas a solid. Yield: 30 mg.

¹HNMR (CDCl₃) δ (ppm): 8.83 (m, 2H); 8.32 (m, 2H); 7.70-7.30 (m, 7H);7.10 (d, 1H); 4.00 (s, 3H).

MS: 459.3 (MH⁺).

HPLC: 99%

EXAMPLE 12 Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (12))

(a) Preparation of methyl 5-bromopicolinate 2

To a 100 mL round bottle flask with 5-bromopicolinic acid (7.0 g, 35mmol) in methanol (80 mL) was added dropwise thionyl chloride (3.0 mL)at ambient temperature. After addition the reaction mixture was heatedto reflux for 3 h. Methanol was removed and ethyl acetate (100 mL) wasadded to the residue and was adjusted pH to 7.0 by addition of sodiumbicarbonate solution. The organic phase was separated and dried oversodium sulfate. The organic solvent was removed and methyl5-bromopicolinate 2 was obtained as white solid which was used for thenext step of the reaction without further purification. Yield: 6.57 g,86.9%

(b) Preparation of methyl 5-(methylthio)picolinate 3

A solution of methyl 5-bromopicolinate 2 (6.20 g, 28.7 mmol) inanhydrous THF (150 mL) was added to sodium methythionide (2.51 g, 35.9mmol) and the reaction was heated to reflux overnight. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1). Methyl 5-(methylthio)picolinate 3 asa white solid was isolated. Yield: 3.0 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.58 (d, 1H), 8.02 (d, 1H), 7.60 (dd,1H), 3.99 (s, 3H), 2.58 (s, 3H).

(c) Preparation of methyl 5-(methylsulfonyl)picolinate 4

To a 100 mL round bottle flask with methyl 5-(methylthio)picolinate 3(1.50 g, 8.2 mmol) in 30 mL of DCM was added mCPBA (5.51 g, 77%, 25.0mmol). The reaction was kept at ambient temperature overnight. Aftersolvent was removed, the residue was purified by column chromatography(eluting with hexane and ethyl acetate 1:1) to give methyl5-(methylsulfonyl)picolinate 4 as a white solid. Yield: 1.35 g, 76.5%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.22 (d, 1H), 8.40 (dd, 1H), 8.36 (d,1H), 4.02 (s, 3H), 3.10 (s, 3H).

(d) Preparation of N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5

2-Chloroaniline (1.20 g, 0.0093 mol) in toluene (20 mL) was added totrimethylaluminum (2.0 M 4.6 mL) then methyl5-(methylsulfonyl)picolinate 4 (1.0 g, 4.65 mol) was added and themixture was heated to 80-90° C. for 2 h. The reaction mixture was cooleddown and 1N HCl solution (10 mL) was added to be acidic. Dichloromethane(100 mL) was then added and the organic phase was further washed withwater (100 mL) and dried over sodium sulfate. The solvent was removedand the residue was mixed with ether (50 mL) and stirred for 0.5 h. Thesolid was filtered and dried to giveN-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 as a light yellowsolid. Yield: 1.26 g, 87.2%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 9.21 (m, 1H), 8.62 (dd,1H), 8.51 (dd, 1H), 8.46 (dd, 1H), 7.45 (dd, 1H), 7.34 (td, 1H), 7.12(td, 1H), 3.20 (s, 3H).

(e) Preparation ofN′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7

N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 (1.26 g, 0.0040 mol)in anhydrous benzene (20 mL) was added to PCl₅ (1.26 g, 6.0 mmol) andthe mixture was heated to reflux overnight. The solvent was removed andthe residue was further dried under high vacuum. CrudeN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasobtained as a yellow solid (1.60 g). TheN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasdissolved into THF anhydrous (30 mL) and the reaction was cooled down to0° C. and hydrazine monohydrate (9.0 mL) was added. The reaction waskept at 0° C. for 10 min and warmed to ambient temperature in 0.5 h. Thesolvent was removed and the residue was purified by columnchromatography (eluting with hexane and ethyl acetate 1:1) to giveN′-(2-chlorophenyl)-5-(methylsulfonyl) picolinimidohydrazide 7 as ayellow solid. Yield: 1.22 g, 92.0%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.02 (dd, 1H), 8.25 (dd, 1H), 8.17 (dd,1H) 7.44 (s, 1H), 7.37 (dd, 1H), 7.16 (td, 1H), 6.89 (td, 1H), 6.45 (dd,1H), 5.62 (s, 2H), 3.06 (s, 3H).

(f) Preparation of3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8

To a 100 mL flask with N′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7 (0.63 g, 1.94 mol) in anhydrous toluene (20 mL)was added maleic anhydride (0.20 g, 2.04 mol) and the reaction was keptat ambient temperature for 1 h and then heated to reflux for 3 h. Aftersolvent was removed, the residue was dried under high vacuum to give3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8 as a yellow solid. Yield: 0.75 g, 95.6%.

(g) Preparation of 2-methoxypyridine-4-carbohydrazide 9

To a solution of 2-methoxypyridine-4-carboxylic acid (2.0 g, 13 mmol) in30 mL of MeOH was added 3 mL of thionyl chloride. The mixture wasstirred at reflux for 2 h. After removal of the solvent, the residue wasdissolved in 30 mL of THF and added to 3 mL of H₂NNH₂.H₂O. The mixturewas heated at reflux for 0.5 h. After removal of solvent, the residuewas purified by column (5-10% of methanol in dichloromethane) to give2-methoxypyridine-4-carbohydrazide 9 as a solid. Yield: 1.76 g, 81%.

¹HNMR (DMSO-d₆) δ (ppm): 9.98 (s, 1H); 8.25 (d, 1H); 7.30 (d, 1H); 7.13(s, 1H); 4.57 (s, 2H); 3.86 (s, 3H).

(h) Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (12))

To a 100 mL flask with3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8 (0.21 g, 0.52 mmol) in dichloromethane (20 mL) was added oxalylchloride (0.20 g, 1.55 mmol) and a drop of DMF. The reaction was kept atambient temperature for 3 h. The solvent was removed and the residue wasfurther dried under high vacuum and cooled to −20° C. Dichloromethane(10 mL) was added, followed by 2-methoxyisonicotinohydrazide 9 (0.10 g,0.6 mmol) and triethylamine (0.5 mL). The reaction mixture was kept atthis temperature for 0.5 h and then warmed to ambient temperature for 1h. The solvent was removed and the crudeN′-[(E)-3-[4-(2-chlorophenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]-2-methoxy-pyridine-4-carbohydrazide10 was used for the next step.

N′-[(E)-3-[4-(2-chlorophenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]-2-methoxy-pyridine-4-carbohydrazide10 was dissolved into dichloromethane (10 mL), and triphenyl phosphine(0.27 g, 1.0 mmol), carbon tetrabromide (0.34 g, 1.04 mmol) andtriethylamine (0.18 mL, 1.30 mmol) were added. The reaction mixture waskept at ambient temperature for 2 h. The final compound was purified bycolumn chromatography (eluting with hexane and ethyl acetate 1:3) togive2-(4-(2-chlorophenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridineas a yellow solid. Yield: 0.031 g, 11%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.75 (d, 1H), 8.62 (d, 1H), 8.35-8.29(m, 2H), 7.70-7.60 (m, 2H), 7.55-7.48 (m, 2H), 7.45-7.39 (m, 1H),7.33-7.32 (m, 1H), 7.12 (d, 1H), 4.01 (s, 3H), 3.09 (s, 3H).

MS: 536.3 (MH⁺).

HPLC: 96%

EXAMPLE 13 Preparation of(E)-4-(5-(2-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)benzonitrile(Compound (10))

(a) Preparation of methyl 5-bromopicolinate 2

To a 100 mL round bottle flask with 5-bromopicolinic acid (7.0 g, 35mmol) in methanol (80 mL) was added dropwise thionyl chloride (3.0 mL)at ambient temperature. After addition the reaction mixture was heatedto reflux for 3 h. Methanol was removed and ethyl acetate (100 mL) wasadded to the residue and was adjusted pH to 7.0 by addition of sodiumbicarbonate solution. The organic phase was separated and dried oversodium sulfate. The organic solvent was removed and methyl5-bromopicolinate 2 was obtained as a white solid which was used for thenext step reaction without further purification. Yield: 6.57 g, 86.9%.

(b) Preparation of methyl 5-(methylthio)picolinate 3

A solution of methyl 5-bromopicolinate 2 (6.20 g, 28.7 mmol) inanhydrous THF (150 mL) was added to sodium methythionide (2.51 g, 35.9mmol) and the reaction was heated to reflux overnight. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1). Methyl 5-(methylthio)picolinate 3 asa white solid was isolated. Yield: 3.0 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.58 (d, 1H), 8.02 (d, 1H), 7.60 (dd,1H), 3.99 (s, 3H), 2.58 (s, 3H).

(c) Preparation of methyl 5-(methylsulfonyl)picolinate 4

To a 100 mL round bottle flask with methyl 5-(methylthio)picolinate 3(1.50 g, 8.2 mmol) in 30 mL of DCM was added mCPBA (5.51 g, 77%, 25.0mmol). The reaction was kept at ambient temperature overnight. After thesolvent was removed, the residue was purified by column chromatography(eluting with hexane and ethyl acetate 1:1) to give methyl5-(methylsulfonyl)picolinate 4 as a white solid. Yield: 1.35 g, 76.5%.

¹H-NMR (300 Hz, CDCl₃) δ ppm 9.22 (d, 1H), 8.40 (dd, 1H), 8.36 (d, 1H),4.02 (s, 3H), 3.10 (s, 3H).

(d) Preparation of N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5

To 2-Chloroaniline (1.20 g, 0.0093 mol) in toluene (20 mL) was addedtrimethylaluminum (2.0 M 4.6 mL) then methyl5-(methylsulfonyl)picolinate 4 (1.0 g, 4.65 mol) was added and themixture was heated to 80-90° C. for 2 h. The reaction was cooled downand 1N HCl solution (10 mL) was added to be acidic. Dichloromethane (100mL) was then added and the organic phase was further washed with water(100 mL) and dried over sodium sulfate. The solvent was removed and theresidue was mixed with ether (50 mL) and stirred for 0.5 h. The solidwas filtered and dried to giveN-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 as a light yellowsolid. Yield: 1.26 g, 87.2%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 9.21 (m, 1H), 8.62 (dd,1H), 8.51 (dd, 1H), 8.46 (dd, 1H), 7.45 (dd, 1H), 7.34 (td, 1H), 7.12(td, 1H), 3.20 (s, 3H).

(e) Preparation ofN′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7

N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 (1.26 g, 0.0040 mol)in anhydrous benzene (20 mL) was added to PCl₅ (1.26 g, 6.0 mmol) andthe mixture was heated to reflux overnight. The solvent was removed andthe residue was further dried under high vacuum. CrudeN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasobtained as a yellow solid (1.60 g). TheN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasdissolved into anhydrous THF (30 mL) and the reaction was cooled down to0° C. and hydrazine monohydrate (9.0 mL) was added. The reaction waskept at 0° C. for 10 min and warmed to ambient temperature in 0.5 h. Thesolvent was removed and the residue was purified by columnchromatography (eluting with hexane and ethyl acetate 1:1) to giveN′-(2-chlorophenyl)-5-(methylsulfonyl) picolinimidohydrazide 7 as ayellow solid. Yield: 1.22 g, 92.0%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.02 (dd, 1H), 8.25 (dd, 1H), 8.17 (dd,1H) 7.44 (s, 1H), 7.37 (dd, 1H), 7.16 (td, 1H), 6.89 (td, 1H), 6.45 (dd,1H), 5.62 (s, 2H), 3.06 (s, 3H).

(f) Preparation of3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylic acid 8

To a 100 mL flask with N′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7 (0.63 g, 1.94 mol) in anhydrous toluene (20 mL)was added maleic anhydride (0.20 g, 2.04 mol) and the reaction was keptat ambient temperature for 1 h and then heated to reflux for 3 h. Aftersolvent was removed and the residue was dried under high vacuum to give3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8 as a yellow solid. Yield: 0.75 g, 95.6%.

(g) Preparation of 4-cyano-benzoic acid hydrazide 9

To a solution of 4-cyano-benzoic acid methyl ester (1.0 g, 13 mmol) in30 mL of MeOH was added 3 mL of H₂NNH₂—H₂O. The mixture was stirred atambient temperature for 16 h. The solid was collected and washed withCH₃OH to give 4-cyano-benzoic acid hydrazide 9 as a solid. Yield: 0.8 g,80%.

¹H NMR (DMSO-d₆) δ (ppm): 7.93 (d, 4H); 4.70 (s, 1H); 3.30 (s, 2H).

(h) Preparation of(E)-4-(5-(2-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)benzonitrile(Compound (10))

To a 100 mL flask with compound 8 (0.21 g, 0.52 mmol) in dichlormethane(20 mL) was added oxalyl chloride (0.20 g, 1.55 mmol) and a drop of DMF.The reaction was kept at ambient temperature for 3 h. The solvent wasremoved and the residue was further dried under high vacuum and cooledto −20° C. Dichloromethane (10 mL) was added, followed by4-cyanobenzohydrazide 9 (0.10 mg, 0.62 mmol) and triethylamine (0.5 mL).The reaction was kept at this temperature for 0.5 h and then roomtemperature for 1 h. The solvent was removed and the crude(E)-N′-(3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acryloyl)-4-cyanobenzohydrazide10 was used for the next step.

(E)-N′-(3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acryloyl)-4-cyanobenzohydrazide10 was dissolved into dichloromethane (10 mL) and triphenyl phosphine(0.27 g, 1.0 mmol), carbon tetrabromide (0.34 g, 1.04 mmol) andtriethylamine (0.18 mL, 1.30 mmol) was added. The reaction was kept atambient temperature for 2 h. The final compound was purified by columnchromatography (eluting with hexane and ethyl acetate 1:3) to give(E)-4-(5-(2-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)vinyl)-1,3,4-oxadiazol-2-yl)benzonitrileas a yellow solid. Yield: 0.035 g, 12.7%.

¹H-NMR (300 Hz, CDCl₃) δ ppm 8.74 (d, 1H), 8.60 (d, 1H), 8.32 (dd, 1H),8.17 (d, 2H), 7.82 (d, 2H), 7.63 (d, 1H), 7.62 (m, 2H), 7.49 (td, 1H),7.41 (dd, 1H), 7.13 (d, 1H), 3.06 (s, 3H).

MS: 530.3 (M+H), 552.20 (M+Na).

HPLC: 98%.

EXAMPLE 14 Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (14))

(a) Preparation of methyl 5-bromopicolinate 2

To a 100 mL round bottle flask with 5-bromopicolinic acid (7.0 g, 0.035mol) in methanol (80 mL) was added thionyl chloride (3.0 mL) dropwise atambient temperature. After the addition the reaction mixture was heatedto reflux for 3 h. Methanol was removed and ethyl acetate (100 mL) wasadded to the residue and was adjusted pH to 7.0 by addition of sodiumbicarbonate solution. The organic phase was separated and dried oversodium sulfate. The organic solvent was removed and methyl5-bromopicolinate 2 was obtained as white solid which was used for thenext step reaction without further purification. Yield: 6.57 g, 86.9%.

(b) Preparation of methyl 5-(methylthio)picolinate 3

A solution of methyl 5-bromopicolinate 2 (6.20 g, 0.0287 mol) inanhydrous THF (150 mL) was added to sodium methythionide (2.51 g, 0.0359mol), and the reaction was heated to reflux overnight. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1). Methyl 5-(methylthio)picolinate 3 asa white solid was isolated. Yield: 3.0 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.58 (d, 1H), 8.02 (d, 1H), 7.60 (dd,1H), 3.99 (s, 3H), 2.58 (s, 3H).

(c) Preparation of methyl 5-(methylsulfonyl)picolinate 4

To a 100 mL round bottle flask with methyl 5-(methylthio)picolinate 3(1.50 g, 0.0082 mol) in 30 mL of DCM was added mCPBA (5.51 g, 77%, 0.025mol). The reaction was kept at ambient temperature overnight. Aftersolvent was removed, the residue was purified by column chromatography(eluting with hexane and ethyl acetate 1:1) to give methyl5-(methylsulfonyl)picolinate 4 as a white solid. Yield: 1.35 g, 76.5%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.22 (d, 1H), 8.40 (dd, 1H), 8.36 (d,1H), 4.02 (s, 3H), 3.10 (s, 3H).

(d) Preparation of N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5

2-Chloroaniline (1.20 g, 0.0093 mol) in toluene (20 mL) was added totrimethylaluminum (2.0 M 4.6 mL) then methyl5-(methylsulfonyl)picolinate 4 (1.0 g, 0.00465 mol) was added and themixture was heated to 80-90° C. for 2 h. The reaction was cooled downand 1N HCl solution (10 mL) was added to be acidic. Dichloromethane (100mL) was then added and the organic phase was further washed with water(100 mL) and dried over sodium sulfate. The solvent was removed and theresidue was mixed with ether (50 mL) and stirred for 0.5 h. The solidwas filtered and dried to giveN-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 as a light yellowsolid. Yield: 1.26 g, 87.2%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 9.21 (m, 1H), 8.62 (dd,1H), 8.51 (dd, 1H), 8.46 (dd, 1H), 7.45 (dd, 1H), 7.34 (td, 1H), 7.12(td, 1H), 3.20 (s, 3H).

(e) Preparation ofN′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7

N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 (1.26 g, 0.0040 mol)in anhydrous benzene (20 mL) was added to PCl₅ (1.26 g, 0.0060 mol) andthe mixture was heated to reflux overnight. The solvent was removed andthe residue was further dried under high vacuum. CrudeN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasobtained as a yellow solid (1.60 g). The compound 6 was dissolved intoanhydrous THF (30 mL) and the reaction was cooled down to 0° C. andhydrazine hydrate (9.0 mL) was added. The reaction was kept at 0° C. for10 min and warmed to ambient temperature in 0.5 h. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1) to giveN′-(2-chlorophenyl)-5-(methylsulfonyl) picolinimidohydrazide 7 as ayellow solid. Yield: 1.22 g, 92.0%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.02 (dd, 1H), 8.25 (dd, 1H), 8.17 (dd,1H) 7.44 (s, 1H), 7.37 (dd, 1H), 7.16 (td, 1H), 6.89 (td, 1H), 6.45 (dd,1H), 5.62 (s, 2H), 3.06 (s, 3H).

(f) Preparation of3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylic acid 8

To a 100 mL flask with N′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7 (0.63 g, 0.00194 mol) in anhydrous toluene (20ml) was added maleic anhydride (0.20 g, 0.00204 mol) and the reactionwas kept at ambient temperature for 1 h and then heated to reflux for 3h. After solvent was removed and the residue was dried under high vacuumto give3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8 as a yellow solid. Yield: 0.75 g, 95.6%.

(g) Preparation of2-(4-(2-chlorophenyl)-5-((E)-2-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (14))

To a 100 mL flask with3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 8 (0.21 g, 0.52 mmol) in dichlormethane (20 mL) was added oxalylchloride (0.20 g, 1.55 mmol) and a drop of DMF. The reaction was kept atambient temperature for 3 h. The solvent was removed and the residue wasfurther dried under high vacuum. Dichloromethane (10 mL) was added,followed by 4-chlorobenzohydrazide 9 (0.10 g, 0.6 mmol) andtriethylamine (0.5 mL). The reaction was kept at this temperature for0.5 h and then ambient temperature for 1 h. The solvent was removed andthe crude4-chloro-N′-[(E)-3-[4-(2-chlorophenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]benzohydrazide10 was used for the next step.

4-chloro-N′-[(E)-3-[4-(2-chlorophenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]benzohydrazide10 was dissolved into dichloromethane (10 mL) and triphenyl phosphine(0.27 g, 1.0 mmol), carbon tetrabromide (0.34 g, 1.04 mmol) andtriethylamine (0.18 mL, 1.30 mmol) was added. The reaction was kept atroom temperature for 2 h. The final compound was purified by columnchromatography (eluting with hexane and ethyl acetate 1:3) to give2-(4-(2-chlorophenyl)-5-((E)-2-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridineas a yellow solid. Yield: 0.03 g, 11%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.74 (d, 1H), 8.60 (d, 1H), 8.32 (dd,1H), 8.17 (d, 2H), 7.82 (d, 2H), 7.63 (d, 1H), 7.62 (m, 2H), 7.49 (d,1H), 7.41 (dd, 1H), 7.13 (d, 1H), 3.06 (s, 3H).

MS: 539.2 (MH⁺).

HPLC: 98%

EXAMPLE 15 Preparation of4-{5-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-ylethynyl]-[1,3,4]oxadiazol-2-yl}-benzonitrile(Compound (33))

(a) Preparation of4-(5-{1,2-Dibromo-2-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-ethyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile2

To a solution of4-(5-{-2-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-vinyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile1 (0.1 g, 0.19 mmol) in DCM (10 mL) was added a solution of Br₂ (0.05 g,0.28 mmol) in DCM (10 mL) dropwise at 0° C. The mixture was stirred atambient temperature for 4 h, washed with NaHSO₃(aq) and H₂O, dried overNa₂SO₄ and concentrated. The residue was subjected to FCC (50-100% ethylacetate/hexanes) to afford4-(5-{1,2-Dibromo-2-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-ethyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile2. Yield: 0.1 g, 77%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.78 (d, 1H), 8.62 (d, 1H), 8.38 (d,1H), 8.20 (d, 2H), 7.80 (d, 2H), 7.72-7.38 (m, 4H), 6.62 (d, 1H), 5.30(d, 1H), 3.09 (s, 3H).

(b) Preparation of4-{5-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-ylethynyl]-[1,3,4]oxadiazol-2-yl}-benzonitrile(Compound (33))

To a solution of4-(5-{1,2-Dibromo-2-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-ethyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile2 (0.1 g, 0.19 mmol) in benzene (10 mL) was added tBuOK (0.05 g, 0.28mmol) portionwise at 0° C. The mixture was stirred at 0° C. for 1 h,treated with diluted acetic acid and ethyl acetate, washed with H₂O,dried over Na₂SO₄ and concentrated. The residue was subjected to FCC(50-100% ethyl acetate/hexanes) to afford4-{5-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-ylethynyl]-[1,3,4]oxadiazol-2-yl}-benzonitrile.Yield: 0.03 g, 39%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.78 (s, 1H), 8.62 (d, 1H), 8.38 (d,1H), 8.20 (d, 2H), 7.82 (d, 2H), 7.62-7.48 (m, 4H), 3.09 (s, 3H).

MS: 528.2 (MH⁺).

HPLC: 94%

EXAMPLE 16 Preparation of2-(4-(2-chloro-4-trifluoromethoxyphenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (56))

(a) Preparation of 4-Cyano-benzoic acid hydrazide 2

To a solution of 4-Cyano-benzoic acid methyl ester 1 (1.0 g, 13 mmol) in30 mL of MeOH was added 3 mL of H₂NNH₂.H₂O. The mixture was stirred atambient temperature for 16 h. The solid was collected and washed withCH₃OH to give 4-Cyano-benzoic acid hydrazide 2 as a solid. Yield: 0.8 g,80%.

¹HNMR (DMSO-d₆) δ (ppm): 7.93 (d, 4H); 4.70 (s, 1H); 3.30 (s, 2H).

(b) Preparation of methyl 5-bromopicolinate 4

To a 100 mL round bottle flask with 5-bromopicolinic acid 3 (7.0 g, 35mmol) in methanol (80 mL) was added dropwise thionyl chloride (3.0 mL)at ambient temperature. After addition the reaction mixture was heatedto reflux for 3 h. Methanol was removed and ethyl acetate (100 mL) wasadded to the residue and was adjusted pH to 7.0 by addition of sodiumbicarbonate solution. The organic phase was separated and dried oversodium sulfate. The organic solvent was removed and methyl5-bromopicolinate 4 was obtained as a white solid which was used fornext step without further purification. Yield: 6.57 g, 86.9%.

(c) Preparation of methyl 5-(methylthio)picolinate 5

A solution of methyl 5-bromopicolinate 4 (6.20 g, 28.7 mmol) inanhydrous THF (150 mL) was added to sodium methythionide (2.51 g, 35.9mmol) and the reaction was heated to reflux overnight. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1). Methyl 5-(methylthio)picolinate 5 asa white solid was isolated. Yield: 3.0 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ ppm 8.58 (d, 1H), 8.02 (d, 1H), 7.60 (dd, 1H),3.99 (s, 3H), 2.58 (s, 3H).

(d) Preparation of methyl 5-(methylsulfonyl)picolinate 6

To a 100 mL round bottle flask with methyl 5-(methylthio)picolinate 5(1.50 g, 8.2 mmol) in 30 mL of DCM was added mCPBA (5.51 g, 77%, 25.0mmol). The reaction was kept at ambient temperature overnight. Aftersolvent was removed, the residue was purified by column chromatography(eluting with hexane and ethyl acetate 1:1) to give methyl5-(methylsulfonyl)picolinate 6 as a white solid. Yield: 1.35 g, 76.5%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.22 (d, 1H), 8.40 (dd, 1H), 8.36 (d,1H), 4.02 (s, 3H), 3.10 (s, 3H).

(e) Preparation of 5-Methanesulfonyl-pyridine-2-carboxylic acid(2-chloro-4-trifluoromethoxy-phenyl)-amide 7

2-Chloro-4-trifluoromethoxy-phenylamine (2.0 g, 9.29 mmol) in toluene(50 mL) was added to trimethylaluminum (2.0 M 4.65 mL) then methyl5-(methylsulfonyl)picolinate 6 (1.0 g, 4.65 mol) was added and themixture was heated to 80-90° C. for 2 h. The reaction was cooled downand 1N HCl solution (10 mL) was added to be acidic. Dichloromethane (100mL) was then added and the organic phase was further washed with water(100 mL) and dried over sodium sulfate. The solvent was removed and theresidue was mixed with ether (50 mL) and stirred for 0.5 h. The solidwas filtered and dried to give 5-methanesulfonyl-pyridine-2-carboxylicacid (2-chloro-4-trifluoromethoxy-phenyl)-amide 7 as a light yellowsolid. Yield: 1.26 g, 65%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 9.21 (m, 1H), 8.68 (dd,1H), 8.52 (dd, 1H), 7.40 (dd, 1H), 7.22 (m, 2H), 3.20 (s, 3H).

(f) Preparation ofN′-(2-chloro-4-trifluoromethoxyphenyl)-5-(methylsulfonyl)picolinimidohydrazide8

5-Methanesulfonyl-pyridine-2-carboxylic acid(2-chloro-4-trifluoromethoxy-phenyl)-amide 7 (1.1 g, 2.79 mmol) inbenzene (20 mL) was added to PCl₅ (1.26 g, 6.0 mmol) and the mixture washeated to reflux overnight. The solvent was removed and the residue wasfurther dried under high vacuum. CrudeN-(2-chloro-4-trifluoromethoxyphenyl)-5-(methylsulfonyl)picolinimidoylchloride was obtained as a yellow solid (1.60 g). TheN-(2-chlor-4-trifluoromethoxyophenyl)-5-(methylsulfonyl)picolinimidoylchloride was dissolved into anhydrous THF (30 mL) and the reaction wascooled down to 0° C. and hydrazine monohydrate (9.0 mL) was added. Thereaction was kept at 0° C. for 10 min and warmed to ambient temperaturein 0.5 h. The solvent was removed and the residue was purified by columnchromatography (eluting with hexane and ethyl acetate 1:1) to giveN′-(2-chloro-4-trifluoromethoxyphenyl)-5-(methylsulfonyl)picolinimidohydrazide8 as a yellow solid. Yield: 0.56 g, 50%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.02 (dd, 1H), 8.25 (dd, 2H), 7.44 (s,1H), 7.32 (dd, 1H), 7.06 (td, 1H), 6.41 (td, 1H), 5.62 (s, 2H), 3.08 (s,3H).

(g) Preparation of3-(4-(2-chlorophenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 9

To a 100 mL flask withN′-(2-chloro-4-trifluoromethoxyphenyl)-5-(methylsulfonyl)picolinimidohydrazide8 (0.45 g, 1.1 mmol) in anhydrous toluene (20 mL) was added maleicanhydride (0.20 g, 2.04 mol) and the reaction was kept at ambienttemperature for 1 h and then heated to reflux for 3 h. After solvent wasremoved, the residue was dried under high vacuum to give3-(4-(2-chloro-4-trifluoromethoxyphenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 9 as a yellow solid which was used for the next step withoutfurther purification. Yield: 0.75 g, 95.6%.

(h) Preparation of2-(4-(2-chloro-4-trifluoromethoxyphenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridine(Compound (56))

To a 100 mL flask with3-(4-(2-chloro-4-trifluoromethoxyphenyl)-5-(5-(methylsulfonyl)pyridin-2-yl)-4H-1,2,4-triazol-3-yl)acrylicacid 9 (0.18 g, 0.368 mmol) in dichlormethane (20 mL) was added oxalylchloride (0.20 g, 1.55 mmol) and a drop of DMF. The reaction was kept atambient temperature for 3 h. The solvent was removed and the residue wasfurther dried under high vacuum and cooled to −20° C. Dichloromethane(10 mL) was added, followed by 4-Cyano-benzoic acid hydrazide 2 (0.18 g,1.1 mmol) and triethylamine (0.5 mL). The reaction mixture was kept atthis temperature for 0.5 h and then ambient temperature for 1 h. Thesolvent was removed and the crudeN′-[(E)-3-[4-(2-chloro-4-trifluoromethoxyphenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]-2-methoxy-pyridine-4-carbohydrazidewas used for the next step.

N′-[(E)-3-[4-(2-chloro-4-trifluoromethoxyphenyl)-5-(5-methylsulfonyl-2-pyridyl)-1,2,4-triazol-3-yl]prop-2-enoyl]-2-methoxy-pyridine-4-carbohydrazidewas dissolved into dichloromethane (10 mL), and triphenylphosphine (0.19g, 0.736 mmol), carbon tetrabromide (0.24 g, 0.736 mmol) andtriethylamine (0.18 mL, 1.30 mmol) was added. The reaction mixture waskept at ambient temperature for 2 h. The final compound was purified bycolumn chromatography (eluting with hexane and ethyl acetate 1:3) togive2-(4-(2-chloro-4-trifluoromethoxyphenyl)-5-((E)-2-(5-(2-methoxypyridin-4-yl)-1,3,4-oxadiazol-2-yl)vinyl)-4H-1,2,4-triazol-3-yl)-5-(methylsulfonyl)pyridineas a yellow solid. Yield: 0.07 g, 30%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.78 (d, 1H), 8.62 (d, 1H), 8.20 (d,2H), 7.78-7.62 (m, 3H), 7.58-7.32 (m, 4H), 7.12 (d, 1H), 3.09 (s, 3H).

MS: 614.2 (MH⁺).

HPLC: 96%

EXAMPLE 17 Preparation of4-[5-(2-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-vinyl)-[1,3,4]oxadiazol-2-yl]-benzonitrile(Compound (57))

(a) Preparation of 5-Hydroxy-pyridine-2-carboxylic acid methyl ester 2

To a 100 mL round bottle flask with 5-Hydroxy-pyridine-2-carboxylic acid1 (1.0 g, 7.2 mmol) in methanol (50 mL) was added dropwise thionylchloride (3.0 mL) at ambient temperature. After addition the reactionmixture was heated to reflux for 3 h. Methanol was removed to give5-Hydroxy-pyridine-2-carboxylic acid methyl ester 2 as a white solidwhich was used for the next step without further purification. Yield:0.957 g, 86.9%.

(b) Preparation of 5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acidmethyl ester 3

A mixture of 5-Hydroxy-pyridine-2-carboxylic acid methyl ester 2 (0.5 g,3.26 mmol), methanesulfonic acid 2,2,2-trifluoro-ethyl ester (1.0 g,5.34 mmol) and K₂CO₃ (1.4 g, 9.8 mmol) in acetonitrile (50 mL) wasstirred overnight. The solid was filtered off and the filtrate wasconcentrated. The residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1) to afford5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acid methyl ester 3 asa white solid. Yield: 0.437 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.48 (d, 1H), 8.16 (d, 1H), 7.38 (dd,1H), 4.48 (q, 2H), 3.99 (s, 3H).

(c) Preparation of 5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acid(2-chloro-phenyl)-amide 4

2-Chloro-4-trifluoromethoxy-phenylamine (2.0 g, 9.29 mmol) in toluene(50 mL) was added to trimethylaluminum (2.0 M 4.65 mL) then5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acid methyl ester 3(1.1 g, 4.65 mol) was added and the mixture was heated to 80-90° C. for2 h. The reaction was cooled down and 1N HCl solution (10 mL) was addedto be acidic.

Dichloromethane (100 mL) was then added and the organic phase wasfurther washed with water (100 mL) and dried over sodium sulfate. Thesolvent was removed and the residue was mixed with ether (50 mL) andstirred for 0.5 h. The solid was filtered and dried to give5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acid(2-chloro-phenyl)-amide 4 as a solid. Yield: 1.0 g, 71%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 8.62 (dd, 1H), 8.40 (dd,1H), 7.32 (dd, 1H), 7.46-7.30 (m, 3H), 7.12-7.08 (m, 1H).

(d) Preparation ofN′-(2-chlorophenyl)-5-(2,2,2-Trifluoro-ethoxy)picolinimidohydrazide 6

To a solution of 5-(2,2,2-Trifluoro-ethoxy)-pyridine-2-carboxylic acid(2-chloro-phenyl)-amide 4 (1.0 g, 3 mmol) in benzene (20 mL) was addedPCl₅ (1.26 g, 6.0 mmol) and the mixture was heated to reflux overnight.The solvent was removed and the residue was further dried under highvacuum. The crudeN-(2-Chloro-phenyl)-5-(2,2,2-trifluoro-ethoxy)-pyridine-2-carboximidoylchloride was dissolved into anhydrous THF (20 mL) and the reaction wascooled down to 0° C. and hydrazine monohydrate (5.0 mL) was added. Thereaction was kept at 0° C. for 10 min and warmed to ambient temperaturein 0.5 h. The solvent was removed and the residue was purified by columnchromatography (eluting with hexane and ethyl acetate 1:1) to giveN′-(2-chloro-phenyl)-5-(2,2,2-Trifluoro-ethoxy)picolinimidohydrazide 6as a yellow solid. Yield: 0.56 g, 50%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.25 (dd, 1H), 8.02 (dd, 1H), 7.44 (s,1H), 7.38 (dd, 1H), 7.30 (dd, 1H), 7.18 (td, 1H), 6.82 (td, 1H), 6.52(d, 1H), 5.32 (s, 2H), 4.40 (q, 2H).

(e) Preparation of3-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-acrylicacid 8

To a 100 mL flask withN′-(2-chloro-4-phenyl)-5-(2,2,2-Trifluoro-ethoxy)picolinimidohydrazide 6(0.45 g, 1.1 mmol) in anhydrous toluene (20 mL) was added maleicanhydride (0.20 g, 2.04 mol) and the reaction was kept at ambienttemperature for 1 h and then heated to reflux for 3 h. After solvent wasremoved and the residue was dried under high vacuum to give3-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-acrylicacid 8 as a yellow solid which was used for the next step withoutfurther purification. Yield: 0.75 g, 95.6%.

(f) Preparation of4-[5-(2-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-vinyl)-[1,3,4]oxadiazol-2-yl]-benzonitrile(Compound (57))

To a 100 mL flask with3-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-acrylicacid 8 (0.18 g, 0.368 mmol) in dichloromethane (20 mL) was added oxalylchloride (0.20 g, 1.55 mmol) and a drop of DMF. The reaction was kept atambient temperature for 3 h. The solvent was removed and the residue wasfurther dried under high vacuum and cooled to −20° C. Dichloromethane(10 mL) was added, followed by 4-Cyano-benzoic acid hydrazide (0.18 g,1.1 mmol) and triethylamine (0.5 mL). The reaction mixture was kept atthis temperature for 0.5 h and then ambient temperature for 1 h. Thesolvent was removed and the crude 4-Cyano-benzoic acidN′-(3-{4-(2-chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-acryloyl)-hydrazidewas used for the next step. 4-Cyano-benzoic acidN′-(3-{4-(2-chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-acryloyl)-hydrazidewas dissolved in dichloromethane (10 mL) and triphenylphosphine (0.19 g,0.736 mmol), carbon tetrabromide (0.24 g, 0.736 mmol) and triethylamine(0.18 mL, 1.30 mmol) were added. The reaction mixture was kept atambient temperature for 2 h. The final compound was purified by columnchromatography (eluting with hexane and ethyl acetate 1:3) to give4-[5-(2-{4-(2-Chloro-phenyl)-5-[5-(2,2,2-trifluoro-ethoxy)-pyridin-2-yl]-4H-[1,2,4]triazol-3-yl}-vinyl)-[1,3,4]oxadiazol-2-yl]-benzonitrileas a yellow solid. Yield: 0.07 g, 30%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.38 (dd, 1H), 8.20 (d, 2H), 8.00 (dd,1H), 7.80 (d, 2H), 7.58-7.32 (m, 6H), 7.18 (d, 1H), 4.40 (q, 2H). MS:550.3 (MH⁺). HPLC: 96%

EXAMPLE 18 Preparation of4-(5-{2-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-1,2-difluoro-vinyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile(Compound (58))

(a) Preparation of Methyl 5-bromopicolinate 2

To a 100 mL round bottle flask with 5-bromopicolinic acid (7.0 g, 35mmol) in methanol (60 mL) was added dropwise thionyl chloride (3.0 mL)at ambient temperature. After addition the reaction mixture was heatedto reflux for 3 h. Methanol was removed and ethyl acetate (80 mL) wasadded to the residue and was adjusted pH to 7.0 by addition of sodiumbicarbonate solution. The organic phase was separated and dried oversodium sulfate. The organic solvent was removed and methyl5-bromopicolinate 2 was obtained as a white solid which was used for thenext step of the reaction without further purification. Yield: 6.57 g,86.9%.

(b) Preparation of Methyl 5-(methylthio)picolinate 3

A solution of methyl 5-bromopicolinate 2 (6.20 g, 28.7 mmol) inanhydrous THF (150 mL) was added to sodium methythionide (2.51 g, 35.9mmol), the reaction was heated to reflux overnight. The solvent wasremoved and the residue was purified by column chromatography (elutingwith hexane and ethyl acetate 1:1). Methyl 5-(methylthio)picolinate 3 asa white solid was isolated. Yield: 3.0 g, 57.1%.

¹H-NMR (300 Hz, CDCl₃) δ ppm 8.58 (d, 1H), 8.02 (d, 1H), 7.60 (dd, 1H),3.99 (s, 3H), 2.58 (s, 3H).

(c) Preparation of Methyl 5-(methylsulfonyl)picolinate 4

To a 100 mL round bottle flask with methyl 5-(methylthio)picolinate 3(1.50 g, 8.2 mmol) in 30 mL of DCM was added mCPBA (5.51 g, 77%, 25.0mmol). The reaction was kept at ambient temperature overnight. Aftersolvent was removed, the residue was purified by column chromatography(eluting with hexane and ethyl acetate 1:1) to give methyl5-(methylsulfonyl)picolinate 4 as a white solid. Yield: 1.35 g, 76.5%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.22 (d, 1H), 8.40 (dd, 1H), 8.36 (d,1H), 4.02 (s, 3H), 3.10 (s, 3H).

(d) Preparation of N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5

2-Chloroaniline (1.20 g, 0.0093 mol) in toluene (20 mL) was added totrimethylaluminum (2.0 M 4.6 mL) then methyl5-(methylsulfonyl)picolinate 4 (1.0 g, 4.65 mol) was added and themixture was heated to 80-90° C. for 2 h. The reaction was cooled downand 1N HCl solution (10 mL) was added to be acidic. Dichloromethane (100mL) was then added and the organic phase was further washed with water(100 mL) and dried over sodium sulfate. The solvent was removed and theresidue was mixed with ether (50 mL) and stirred for 0.5 h. The solidwas filtered and dried to giveN-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 as a light yellowsolid. Yield: 1.26 g, 87.2%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 10.6 (s, 1H), 9.21 (m, 1H), 8.62 (dd,1H), 8.51 (dd, 1H), 8.46 (dd, 1H), 7.45 (dd, 1H), 7.34 (td, 1H), 7.12(td, 1H), 3.20 (s, 3H).

(e) Preparation ofN′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7

N-(2-chlorophenyl)-5-(methylsulfonyl)picolinamide 5 (1.26 g, 0.0040 mol)in anhydrous benzene (20 mL) was added to PCl₅ (1.26 g, 6.0 mmol) andthe mixture was heated to reflux overnight. The solvent was removed andthe residue was further dried under high vacuum. CrudeN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasobtained as a yellow solid (1.60 g). TheN-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidoyl chloride 6 wasdissolved into anhydrous THF (30 mL) and the reaction was cooled down to0° C. and hydrazine monohydrate (9.0 mL) was added. The reaction waskept at 0° C. for 10 min and warmed to ambient temperature in 0.5 h. Thesolvent was removed and the residue was purified by columnchromatography (eluting with hexane and ethyl acetate 1:1) to giveN′-(2-chlorophenyl)-5-(methylsulfonyl) picolinimidohydrazide 7 as ayellow solid. Yield: 1.22 g, 92.0%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 9.02 (dd, 1H), 8.25 (dd, 1H), 8.17 (dd,1H) 7.44 (s, 1H), 7.37 (dd, 1H), 7.16 (td, 1H), 6.89 (td, 1H), 6.45 (dd,1H), 5.62 (s, 2H), 3.06 (s, 3H).

(f) Preparation of3-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-2,3-difluoro-acrylicacid 9

To a 100 mL flask withN′-(2-chlorophenyl)-5-(methylsulfonyl)picolinimidohydrazide 7 (0.84 g,2.59 mmol) in anhydrous toluene (20 mL) was added3,4-difluoro-furan-2,5-dione (0.36 g, 2.59 mmol) and the reaction waskept at ambient temperature for 1 h to give(2Z)-3-((Z)—N′-(2-chlorophenyl)-5-(methylsulfonyl)pyridine-2-carboxamidocarbamoyl)-2,3-difluoroacrylicacid 8 and then heated to reflux for 3 h. After solvent was removed theresidue was dried under high vacuum to give3-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-2,3-difluoro-acrylicacid 9 as a yellow solid (1.15 g, 96%) which was used for the next stepwithout further purification.

(g) Preparation of4-(5-{2-[4-(2-Chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-1,2-difluoro-vinyl}-[1,3,4]oxadiazol-2-yl)-benzonitrile(Compound (58))

To a 100 mL flask with3-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-2,3-difluoro-acrylicacid 9 (0.3 g, 0.68 mmol) in dichloromethane (20 mL) was added oxalylchloride (0.40 g, 3.1 mmol) and a drop of DMF. The reaction was kept atambient temperature for 3 h. The solvent was removed and the residue wasfurther dried under high vacuum and cooled to −20° C. Dichloromethane(10 mL) was added, followed by 4-cyano-benzoic acid hydrazide 9 (0.33 g,2.04 mmol) and triethylamine (0.5 mL). The reaction mixture was kept atthis temperature for 0.5 h and then ambient temperature for 1 h. Thesolvent was removed and the crude 4-cyano-benzoic acidN′-{3-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-2,3-difluoro-acryloyl}-hydrazide11 was used for the next step.

4-Cyano-benzoic acidN′-{3-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-2,3-difluoro-acryloyl}-hydrazide11 was dissolved into dichloromethane (10 mL) and triphenylphosphine(0.27 g, 1.02 mmol), carbon tetrabromide (0.34 g, 1.02 mmol) andtriethylamine (0.18 mL, 1.30 mmol) were added. The reaction mixture waskept at ambient temperature for 2 h. The final compound was purified bycolumn chromatography (eluting with methanol/DCM 1/99-5/95) to give4-(5-{2-[4-(2-chloro-phenyl)-5-(5-methanesulfonyl-pyridin-2-yl)-4H-[1,2,4]triazol-3-yl]-1,2-difluoro-vinyl}-[1,3,4]oxadiazol-2-yl)-benzonitrileas a solid. Yield: 0.05 g, 13%.

¹H-NMR (300 Hz, CDCl₃) δ (ppm): 8.77 (s, 1H), 8.61 (d, 1H), 8.38 (d,1H), 8.10 (d, 2H), 7.80 (d, 2H), 7.58-7.51 (m, 3H), 7.46-7.41 (m, 1H),3.10 (s, 3H).

MS: 566.2 (MH⁺).

HPLC: 95%

EXAMPLE 19 IC₅₀ Values of Compounds

IC₅₀ values were calculated using STF/REN HEK293 cells (stablytransfected with ST-Luc (7× TCF binding sites) and Renilla (Promega)plasmids). HEK293 cells (80,000 cells per well) were seeded in 96-wellplates coated with poly-L lysine. 24 hours after seeding, the cells wereincubated for an additional 24 hours with various compoundconcentrations in 50% Wnt3a containing conditioned media (from LWnt3a-expressing cells (ATCC)). A minimum of three replicates was usedfor each sample. After compound exposure, the cells were lysed and thefirefly luciferase and Renilla activities were measured on a GloMax® 96Microplate Luminometer with Dual Injectors (Promega).

XLfit (idbs) was used to determine the IC₅₀ values in inhibitionexperiments. The data were fit to the following formula:

IC₅₀: Langmuir Binding Isotherm:fit=((A+(B*x))+(((C−B)*(1−exp((−1*D)*x)))/D))res=(y−fit)

Table 1 shows the IC₅₀ values of certain compounds. All values shown areaverage values from multiple experiments.

TABLE 1 Compound IC₅₀ ±Standard No. μM Deviation  (1) 0.94 0.01  (2)0.28 0.01  (3) 0.19 0.01  (4) 0.24 0.00  (5) >10  (6) 1.2 0.23  (7) >10 (8) 6.3 0.11 (10) 0.05 0.02 (11) 0.08 0.05 (12) 0.07 0.02 (13) 0.110.04 (14) 0.04 0.01 (33) 3.6 0.84 (56) 0.39 0.14 (57) 0.10 0.13 (58) >10

EXAMPLE 20 Stability of Compounds

Experimental Procedure

Pooled human or rat liver microsomes (pooled male and female) wereprepared and stored at −80° C. prior to use. Microsomes (final proteinconcentration 0.5 mg/mL), 0.1M phosphate buffer (pH 7.4) and the testcompound (final substrate concentration=3 μM; final DMSOconcentration=0.25%) were pre-incubated at 37° C. prior to the additionof NADPH (final concentration=1 mM) to initiate the reaction. The finalincubation volume was 25 μL. A control incubation was included for eachcompound tested in which 0.1M phosphate buffer (pH 7.4) was addedinstead of NADPH (minus NADPH). Two control compounds were included witheach species. All incubations were performed singularly for each testcompound.

Each compound was incubated for 0, 5, 15, 30 and 45 mins. The control(minus NADPH) was incubated for 45 mins only. The reactions were stoppedby the addition of 50 μL methanol containing internal standard at theappropriate time points. The incubation plates were centrifuged at 2,500rpm for 20 mins at 4° C. to precipitate the protein.

Quantitative Analysis

Following protein precipitation, the sample supernatants were combinedin cassettes of up to 4 compounds and analysed using Cyprotex genericLC-MS/MS conditions. Optionally, if metabolite profiling was requestedfollowing the stability assay a second assay was performed in which thecompound was incubated four times and the four resulting incubationswere pooled to yield a higher sample concentration for analysis. Thetime point at which 30-50% of parent had degraded could then beinvestigated at 3 different levels of metabolite profiling and/oridentification.

Data Analysis

From a plot of ln peak area ratio (compound peak area/internal standardpeak area) against time, the gradient of the line is determined.Subsequently, half-life and intrinsic clearance are calculated using theequations below:Elimination rate constant (k)=(−gradient)

${{Half}\mspace{14mu}{life}\mspace{14mu}\left( t_{1/2} \right)\mspace{14mu}({mins})} = \frac{0.693}{k}$${{Intrinsic}\mspace{14mu}{Clearance}\mspace{14mu}\left( {CL}_{int} \right)\mspace{14mu}\left( {{{{µL}/\min}/{mg}}\mspace{14mu}{protein}} \right)} = \frac{V \times 0.693}{t_{1/2}}$

-   -   where V=Incubation volume (μL/mg microsomal protein).

Two control compounds were included in the assay and if the values forthese compounds were not within the specified limits the results wererejected and the experiment repeated.

Results

Compound T ½ human liver No. microsomes (mins) (1) 162 (2) 224 (3) 37.6(4) 44.9 (5) 5.41 (6) 48 (8) 128 (10)  101 (11)  59.6 (12)  37.1 (13) 212 (14)  371

EXAMPLE 21 Inhibition of Cell Growth

The growth of cells was assessed in the presence and absence of compound(10) using a proliferation assay in an InuCyte™ live cell imager.

Experimental Procedure

All cells were purchased from ATCC (American Type Culture Collection)and maintained according to the supplier's recommendations. Cells usedwere RKO, HCT-15, WiDr, HT29, DLD-1, COLO320DM and COLO205 cells. 1,000cells were seeded in 96-well plates with the recommended media. The dayafter seeding, the cell culture medium was exchanged to solutions thatcontained 0.05% DMSO and 5, 1 or 0.1 μmol/L compound (10). All sampleswere assessed in a minimum of six replicates. Plates were incubated inan IncuCyte™ (Essen BioScience) inside a cell culture incubator. Imageswere captured every second hour to monitor cell proliferation.

Results

Mutations in the Adenomatouse Polyposis Coli (APC) gene, which occur inmost colorectal cancers (CRC), lead to ineffective degradation ofβ-catenin and aberrant up-regulation of Wnt signaling. As a result, CRCcells may undergo cell cycle arrest as a result of antagonized canonicalWnt. Therefore, the kinetics of cell proliferation was monitored inselected Wnt responsive CRC cell lines by using an IncuCyte lifetracking system. In parallel, the colorectal cancer cell line RKO, whichcontains wild type APC and β-catenin and exhibits Wnt-independent cellgrowth was used as a control. The cell growth profiles for the celllines RKO, HCT-15, WiDr, HT29, DLD-1, COLO320DM and COLO205, aftertreatment with DMSO (control), 0.1 μmol/L, 1 μmol/L, and 5 μmol/Lcompound (10) are shown in FIGS. 1A to 1G, respectively.

Inhibition of canonical Wnt signaling by compound (10) promotes cellcycle arrest and specifically reduces proliferation in APC mutant CRCcells. Cell growth curves, as measured by IncuCyte™, show aconcentration-dependent decrease of proliferation in various APC mutantCRC cells compared to the Wnt-independent CRC control cell line RKO thatcontains wild type APC. Plots show representative values from a minimumof two independent experiments and all relative standard deviations arebelow 20%. The cell lines HCT-15, WiDr, HT29, DLD-1, COLO320DM andCOLO205 showed dose-dependent growth inhibition, while the control cellline RKO did not. In each case, apart from the control, the curve forDMSO (black line) reaches confluency in a shorter time than the othersamples, which show dose-dependent inhibition of cell growth (i.e.greater inhibition of growth at increasing concentration of compound(10)—0.1 μM, light grey line; 1 μM, mid-grey line; and 5 μM, dark greyline).

EXAMPLE 22 Inhibition of the Wnt Pathway

Experimental Procedure

The inhibitory activities of compound (10) and XAV939 (Novartis; Huanget al., Nature (2009), 461, pp. 614-20), which is used as a positivecontrol, were tested at various doses (in duplicate) usingChemiluminescent Assay Kits (BPS Bioscience, Nordic Biosite) againstTNKS1 (Cat No. 80564), TNSK2 (Cat No. 80566) and PARP1 (Cat No. 80551).The procedures were performed according to the manufacturer's protocols.

Results

To test whether compound (10) decreased canonical Wnt signaling byinhibiting the PARP domain of TNKS1/2, biochemical assays for monitoringthe activity of TNKS1/2 and PARP were performed. Compound (10) decreasedauto-PARsylation of TNKS1 and TNKS2 in vitro with IC₅₀-values of 46nmol/L and 19 nmol/L, respectively. However, in contrast to XAV939,compound (10) exhibited no inhibition of PARP1 at doses up to 20 μmol/L.The results of the Experiments are shown in FIG. 2 (TNKS1), 3 (TNKS2)and 4 (PARP). In each case, the figure A shows the results obtained withXAV939 and figure B shows the results obtained with compound (10).

The IC₅₀ values calculated from the results of this experiment are asfollows:

Assay IC₅₀ value for XAV939 IC₅₀ value for compound (10) TNKS1 15 nM 46nM TNKS2 7.8 nM 19 nM PARP 0.64 μM >20 μM

The results of this experiment indicate that compound (10) blockscanonical Wnt signaling by specifically inhibiting auto-PARsylation ofTNKS1/2 while leaving PARP1 activity unaffected.

EXAMPLE 23 Immunohistochemical Analysis

Experimental Procedure

50,000 SW480 cells (ATCC—maintained according to the supplier'srecommendations) were seeded in 24-well plates on glass slides andexposed to DMSO (control) or 0.5 μmol/L of compound (10) for 18 hours.After incubation, the cells were fixed in 4% PFA in PBS for 10 minutes.

Immunostaining was performed as described in standard protocols. Primaryantibodies used were β-catenin (610153, BD Transduction Laboratories™)or AXIN2 (76G6, Cell Signaling Technology). Secondary antibodies usedwere DyLight549 (555) donkey-anti-mouse and Cy2-donkey-anti-rabbit (bothJackson ImmunoResearch, 1:1000). The samples were imaged using a ZeissAxiovert 200M Fluorescence/Live cell Imaging Microscope at 40 timesmagnification. A Zeiss LSM780 at 63 times magnification was used forconfocal microscopy

Results

To gain an insight into the changes in cellular distribution of AXIN2and β-catenin, SW480 cells treated with compound (10) were analyzed byimmunofluorescence. A general reduction of total β-catenin, accompaniedwith a strong reduction of nuclear β-catenin, was detected at a dose of500 nmol/L in most, but not all, compound (10) treated SW480 cells.Confocal microscopy (equal shutter speeds) revealed that the levels ofcytoplasmic AXIN2 were significantly increased and large protein foci,apparently representing accumulated destruction complexes, were observed(magnified insert in FIG. 5). Degradation of β-catenin after compound(10) exposure appeared to be orchestrated by stabilization of AXIN2 inthe destruction complex. Images of cells treated with DMSO and compound(10) are shown in FIG. 5.

The invention claimed is:
 1. A method for treating a condition ordisease which is affected by over-activation of signaling in the Wntpathway, said method comprising administering to an individual in needthereof a compound of general formula I:

(wherein Z¹ represents

Z² represents phenyl, pyridyl, pyrimidinyl or oxadiazolyl optionallysubstituted by one or more groups R_(a); where each R_(a) may beidentical or different and may be selected from F, Cl, Br, I, C₁₋₆alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, —CN, —NO₂, —OR, —SR,—C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR,—S(O)R, —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R isindependently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆alkynyl); R¹ represents a phenyl or pyridyl group optionally substitutedby one or groups R_(b); where each R_(b) may be identical or differentand may be selected from F, Cl, Br, I, C₁₋₆ alkyl optionally interruptedby one or more —O—, —S— or —NR— groups, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄haloalkyl, —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂,—C(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R, —S(O)₂R, —S(O)OR or—S(O)₂NR₂ group (where each R is independently H, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl); R² represents a phenyl groupoptionally substituted by one or more groups R_(c); where each R_(c) maybe identical or different and may be selected from F, Cl, Br, I, C₁₋₆alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, —CN, —NO₂, —OR, —SR,—C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR,—S(O)R, —S(O)₂R, or —S(O)OR group (where each R is independently H, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl); L¹ represents aC₁₋₄ alkylene group optionally substituted by one or more groups R_(d),wherein one or more methylene groups are each replaced by a groupselected from —CR_(e)═CR_(f)—, —C≡C— and —C═C═C—; and wherein one ormore methylene groups may each additionally be replaced by a group Y¹;where each Y¹ is independently selected from —O—, —S—, —NH—, —NR′″—,—NR′″—C(O)—, —C(O)—NR′″—, —C(O)—, —S(O₂)—, —S(O)— and —CR′″═N— (whereeach R′″ is independently hydrogen or C₁₋₆ alkyl); where each R_(d) maybe identical or different and may be selected from C₁₋₆ alkyl, hydroxy,C₁₋₆ alkoxy, F, Cl, Br and I; and where R_(e) and R_(f) areindependently selected from H, C₁₋₃ alkyl, halogen, C₁₋₃ haloalkyl, —CN,—NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OPO₃R, —OSO₂R and —OSiR₄(where each R is independently H, C₁₋₆ alkyl or C₁₋₆ haloalkyl); L²represents a bond; and L³ represents a bond or a stereoisomer orpharmaceutically acceptable salt thereof, wherein the condition ordisease which is affected by over-activation of signaling in the Wntpathway is a cancer selected from colon cancers, pancreatic cancer,gastric cancer, liver cancers, Wilms tumor of the kidney,medulloblastoma, skin cancers, non-small cell lung cancer, cervicalcancer, ovarian endometrial cancer, bladder cancer, anaplastic thyroidcancer, head and neck cancer, breast cancer, prostate cancer, andglioblastoma.
 2. An in vivo or in vitro method of promoting and/ordirecting cellular differentiation comprising contacting a progenitorcell with an effective amount of a compound of general formula I:

(wherein Z¹ represents

Z² represents phenyl, pyridyl, pyrimidinyl or oxadiazolyl optionallysubstituted by one or more groups R_(a); where each R_(a) may beidentical or different and may be selected from F, Cl, Br, I, C₁₋₆alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, —CN, —NO₂, —OR, —SR,—C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR,—S(O)R, —S(O)₂R, —S(O)OR or —S(O)₂NR₂ group (where each R isindependently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆alkynyl); R¹ represents a phenyl or pyridyl group optionally substitutedby one or groups R_(b); where each R_(b) may be identical or differentand may be selected from F, Cl, Br, I, C₁₋₆ alkyl optionally interruptedby one or more —O—, —S— or —NR— groups, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄haloalkyl, —CN, —NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂,—C(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR, —S(O)R, —S(O)₂R, —S(O)OR or—S(O)₂NR₂ group (where each R is independently H, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl); R² represents a phenyl groupoptionally substituted by one or more groups R_(c); where each R_(c) maybe identical or different and may be selected from F, Cl, Br, I, C₁₋₆alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, —CN, —NO₂, —OR, —SR,—C(O)R, —C(O)OR, —OC(O)R, —OC(O)NR₂, —NR₂, —NR—C(O)R, —NR—C(O)OR,—S(O)R, —S(O)₂R, or —S(O)OR group (where each R is independently H, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl or C₂₋₆ alkynyl); L¹ represents aC₁₋₄ alkylene group optionally substituted by one or more groups R_(d),wherein one or more methylene groups are each replaced by a groupselected from —CR_(e)═CR_(f)—, —C≡C— and —C═C═C—; and wherein one ormore methylene groups may each additionally be replaced by a group Y¹;where each Y¹ is independently selected from —O—, —S—, —NH—, —NR′″—,—NR′″—C(O)—, —C(O)—NR′″—, —C(O)—, —S(O₂)—, —S(O)— and —CR′″═N— (whereeach R′″ is independently hydrogen or C₁₋₆ alkyl); where each R_(d) maybe identical or different and may be selected from C₁₋₆ alkyl, hydroxy,C₁₋₆ alkoxy, F, Cl, Br and I; and where R_(e) and R_(f) areindependently selected from H, C₁₋₃ alkyl, halogen, C₁₋₃ haloalkyl, —CN,—NO₂, —OR, —SR, —C(O)R, —C(O)OR, —OC(O)R, —OPO₃R, —OSO₂R and —OSiR₄(where each R is independently H, C₁₋₆ alkyl or C₁₋₆ haloalkyl); L²represents a bond; and L³ represents a bond or a stereoisomer orpharmaceutically acceptable salt thereof.
 3. The method of claim 1,wherein, in said compound of general formula I, L¹ is a C₁₋₄ alkylenegroup optionally substituted by one or more groups R_(d), wherein one ormore methylene groups are each replaced by a group —CR_(e)═CR_(f)— or bya group —C≡C—; where each R_(d) may be identical or different and may beselected from C₁₋₆ alkyl, hydroxy, C₁₋₆ alkoxy, F, Cl, Br and I; andwhere R_(e) and R_(f) are independently selected from H, C₁₋₃ alkyl,halogen, and —CN.
 4. The method of claim 1, wherein, in said compound ofgeneral formula I, Z² represents phenyl, pyridyl or pyrimidinyloptionally mono- or di-substituted by a group R_(a); where R_(a) may beselected from F, Cl, Br, I, hydroxy, C₁₋₆ alkoxy, and —S(O)₂R (where Ris H or C₁₋₃ alkyl); R¹ represents a phenyl or pyridyl group optionallymono-substituted by group R_(b); where R_(b) is selected from F, Cl, Br,I, C₁₋₆ alkyl, C₁₋₆ alkoxy, hydroxy and —CN; R² represents a phenylgroup optionally mono- or di-substituted by a group R_(c); where eachR_(c) may be identical or different and may be selected from F, Cl, Br,I, C₁₋₆ alkyl, and C₁₋₆ alkoxy; L¹ represents a C₁₋₄ alkylene groupoptionally substituted by one or more groups R_(d), wherein one or twomethylene groups are each replaced by a group —CR_(e)═CR_(f)— or by agroup —C≡C—; where each R_(d) may be identical or different and may beselected from C₁₋₆ alkyl, hydroxy and C₁₋₆ alkoxy; where R_(e) and R_(f)are independently selected from H, C₁₋₃ alkyl and halogen.
 5. The methodof claim 1, wherein, in said compound of general formula I, L¹ is agroup —CR_(e)═CR_(f)—, —C≡C— or —C═C═C— in which R_(e) and R_(f) are asdefined in claim
 1. 6. The method of claim 1, wherein, in said compoundof general formula I, L¹ is a group —CR_(e)═CR_(f)— in which R_(e) andR_(f) are independently selected from H and methyl.
 7. The method ofclaim 1, wherein said compound of general formula I is a compound havingthe formula IIc or IId:

(wherein Z² is an optionally substituted pyridyl, phenyl or pyrimidinylring; R¹ is a substituted phenyl or pyridyl ring; R² is an optionallysubstituted phenyl ring; and L¹ is cis or trans —CH═CH—) or astereoisomer or pharmaceutically acceptable salt thereof.
 8. The methodof claim 1, wherein said compound of general formula I is selected fromthe following: Compound No. Structure  (1)

 (2)

 (3)

 (4)

 (5)

 (6)

 (8)

 (9)

(10)

(11)

(12)

(13)

(14)

(15)

(16)

(17)

(18)

(19)

(21)

(22)

(23)

(24)

(25)

(26)

(27)

(28)

(29)

(31)

(32)

(33)

(34)

(35)

(36)

(37)

(38)

(39)

(40)

(41)

(42)

(43)

(44)

(45)

(46)

(47)

(48)

(49)

(50)

(51)

(52)

(53)

(54)

(55)

or a stereoisomer or pharmaceutically acceptable salt thereof.
 9. Themethod of claim 8, wherein said compound of general formula I isselected from the following: Compound Nos. (1), (2), (3), (4), (5), (6),(8), (10), (11), (12), (14), and (33), their stereoisomers andpharmaceutically acceptable salts thereof.
 10. The method of claim 8,wherein said compound of general formula I is selected from thefollowing: Compound Nos. (2), (3), (4), (8), (10), (11), (12), and (14),their stereoisomers and pharmaceutically acceptable salts thereof. 11.The method of claim 1, wherein said compound of general formula I isprovided in a pharmaceutical formulation which comprises a compound ofgeneral formula I as defined in claim 1, or a pharmaceuticallyacceptable salt thereof, together with one or more pharmaceuticallyacceptable carriers or excipients.
 12. The method of claim 2, wherein,in said compound of general formula I, L¹ is a C₁₋₄ alkylene groupoptionally substituted by one or more groups R_(d), wherein one or moremethylene groups are each replaced by a group —CR_(e)═CR_(f)— or by agroup —C≡C—; where each R_(d) may be identical or different and may beselected from C₁₋₆ alkyl, hydroxy, C₁₋₆ alkoxy, F, Cl, Br and I; andwhere R_(e) and R_(f) are independently selected from H, C₁₋₃ alkyl,halogen, and —CN.
 13. The method of claim 2, wherein, in said compoundof general formula I, Z² represents phenyl, pyridyl or pyrimidinyloptionally mono- or di-substituted by a group R_(a); where R_(a) may beselected from F, Cl, Br, I, hydroxy, C₁₋₆ alkoxy, and —S(O)₂R (where Ris H or C₁₋₃ alkyl); R¹ represents a phenyl or pyridyl group optionallymono-substituted by group R_(b); where R_(b) is selected from F, Cl, Br,I, C₁₋₆ alkyl, C₁₋₆ alkoxy, hydroxy and —CN; R² represents a phenylgroup optionally mono- or di-substituted by a group R_(c); where eachR_(c) may be identical or different and may be selected from F, Cl, Br,I, C₁₋₆ alkyl, and C₁₋₆ alkoxy; L¹ represents a C₁₋₄ alkylene groupoptionally substituted by one or more groups R_(d), wherein one or twomethylene groups are each replaced by a group —CR_(e)═CR_(f)— or by agroup —C≡C—; where each R_(d) may be identical or different and may beselected from C₁₋₆ alkyl, hydroxy and C₁₋₆ alkoxy; where R_(e) and R_(f)are independently selected from H, C₁₋₃ alkyl and halogen.
 14. Themethod of claim 2, wherein, in said compound of general formula I, L¹ isa group —CR_(e)═CR_(f)—, —C≡C— or —C═C═C— in which R_(e) and R_(f) areas defined in claim
 1. 15. The method of claim 2, wherein, in saidcompound of general formula I, L¹ is a group —CR_(e)═CR_(f)— in whichR_(e) and R_(f) are independently selected from H and methyl.
 16. Themethod of claim 2, wherein said compound of general formula I is acompound having the formula IIc or IId:

(wherein Z² is an optionally substituted pyridyl, phenyl or pyrimidinylring; R¹ is a substituted phenyl or pyridyl ring; R² is an optionallysubstituted phenyl ring; and L¹ is cis or trans —CH═CH—) or astereoisomer or pharmaceutically acceptable salt thereof.
 17. The methodof claim 2, wherein said compound of general formula I is selected fromthe following: Compound No. Structure  (1)

 (2)

 (3)

 (4)

 (5)

 (6)

 (8)

 (9)

(10)

(11)

(12)

(13)

(14)

(15)

(16)

(17)

(18)

(19)

(21)

(22)

(23)

(24)

(25)

(26)

(27)

(28)

(29)

(31)

(32)

(33)

(34)

(35)

(36)

(37)

(38)

(39)

(40)

(41)

(42)

(43)

(44)

(45)

(46)

(47)

(48)

(49)

(50)

(51)

(52)

(53)

(54)

(55)

or a stereoisomer or pharmaceutically acceptable salt thereof.
 18. Themethod of claim 17, wherein said compound of general formula I isselected from the following: Compound Nos. (1), (2), (3), (4), (5), (6),(8), (10), (11), (12), (14), and (33), their stereoisomers andpharmaceutically acceptable salts thereof.
 19. The method of claim 17,wherein said compound of general formula I is selected from thefollowing: Compound Nos. (2), (3), (4), (8), (10), (11), (12), and (14),their stereoisomers and pharmaceutically acceptable salts thereof. 20.The method of claim 2, wherein said compound of general formula I isprovided in a pharmaceutical formulation which comprises a compound ofgeneral formula I as defined in claim 1, or a pharmaceuticallyacceptable salt thereof, together with one or more pharmaceuticallyacceptable carriers or excipients.